Bioactive peptides discovery from spirulina classification of common characteristics of antihypertensive peptides Bioactive peptides discovery from spirulina classification of common characteristics of antihypertensive peptides Bioactive peptides discovery from spirulina classification of common characteristics of antihypertensive peptides Bioactive peptides discovery from spirulina classification of common characteristics of antihypertensive peptides
THAI NGUYEN UNIVERSITY UNIVERSITY OF AGRICULTURE AND FORESTRY LUONG TAI CHIEN Topic title: BIOACTIVE PEPTIDES DISCOVERY FROM SPIRULINA: CLASSIFICATION OF COMMON CHARACTERISTICS OF ANTIHYPERTENSIVE PEPTIDES BACHELOR THESIS Study Mode : Full-time Major : Biotechnology Faculty : Biotechnology and Food Technology Batch : 2013 – 2017 Thai Nguyen, 6/2017 THAI NGUYEN UNIVERSITY UNIVERSITY OF AGRICULTURE AND FORESTRY LUONG TAI CHIEN Topic title: BIOACTIVE PEPTIDES DISCOVERY FROM SPIRULINA: CLASSIFICATION OF COMMON CHARACTERISTICS OF ANTIHYPERTENSIVE PEPTIDES BACHELOR THESIS Study Mode : Full-time Major : Biotechnology Faculty :Biotechnology and Food Technology Batch : 2013 – 2017 Supervisors :Asst Prof Dr Marasri Ruengjitchatchawalya Dr Duong Van Cuong Supervisors Supervisor Asst Prof Dr Marasri Ruengjitchatchawalya Dr Duong Van Cuong Thai Nguyen, 6/2017 DOCUMENTATION PAGE WITH ABSTRACT Thai Nguyen University of Agriculture and Forestry Major Biotechnology Student name Luong Tai Chien Student ID DTN1353150004 Bioactive peptides discovery from Spirulina: Classification Thesis title of common characteristics of antihypertensive peptides Asst Prof Dr Marasri Ruengjitchatchawalya Supervisors Dr Duong Van Cuong Abstract: Arthrospira (Spirulina)platensisis a spiral filamentous cyanobacteriumcontaining high protein content about 55-70%as dry weight The alga has been used as human food- and animal feed supplements.Attempt to discover bioactive peptides from A platensis, Algal Biotechnology research group at King Mongkut’s University of Technology Thonburi (KMUTT), Thailand,have investigated an in-silico digestion of the algal proteome and classified the obtained peptides regarding their bioactivities In this report, hands-on experience about the algal, A platensisC1, cultivation; cell harvesting, as well as protein extraction/ separation and a bioactivity assay, antihypertensive activity (AH) of some obtained peptides has been described In addition, using a computational tool, Clustal Omega, to determine common characteristics of the putative anti-hypertensive peptides showed that 10 common amino acidgroups were classified, including major groups of G, P, Y and L The reference Novel AH peptidewas in thegroup Y with the common amino acids―YY‖ Besides, these peptides were mostly contained both aliphatic and aromatic amino acid (A, G, I, L, P, V, F, W, Y) and also hydrophobic amino acid residue (A, I, L, F, V, P, G) Keywords Number of pages Spirulina (Arthrospira) platensis C1, Bioactive peptides, common characteristics 37 i ACKNOWLEDGEMENT This thesis was completed by the support and assistance of a number of people whom I would like to personally thank First and foremost, I would like to express my gratitude to the both my supervisors Asst Prof Dr Marasri Ruengjitchatchawalya of the school of Bioresources and Technology, King Mongkut’s University of Technology Thonburi (KMUTT), Thailand and Dr Duong Van Cuong from the biotechnology and food technology department of Thai Nguyen University of Agriculture and Forestry (TUAF) Whose expertise and understanding guided me through my internship, providing useful advice for the improvement of this work Some special thanks also to my lab facilitator, PhD.Candidate,Miss Krittima Anekthanakul, for sharing her knowledge, formaking experiments look amusing Big thanks also go to every people on Algal Biotechnology Laboratory (AGB research group) and Bioinformatics and Systems Biology (BIF) Program at KMUTT for helping me fit in and feel welcome from the moment and for the unlimited patience to explain me every doubt I had during my internship.I would also like to acknowledge my teachers at TUAF, Dr Duong Van Cuong, MSc Trinh Thi Chung, Dr Nguyen Xuan Vu and MSc Duong Manh Cuong that contributed to making this work and had an enjoyable and fulfilling experience Many thank you and best regards Student Luong Tai Chien ii CONTENTS PART I INTRODUCTION 1.1 Background and motivation 1.1.1 Arthrospira (Spirulina) platensis 1.1.1.1 Classification 1.1.1.2 Morphology of Arthrospira 1.1.1.3 Ultrastructure of A platensis 1.1.1.4 The life cycle of Arthrospira (Spirulina) 1.1.2 Bioactive peptides of A platensis C1 1.1.2.1 Antioxidant and Antiinflammatory activity 1.1.2.2 Antihypertension 1.1.2.3 Antimicrobial activity 10 1.1.2.4 Antidiabetes and Antiobesity 10 1.1.2.5 Anticancer 10 1.1.3 Computational Tool: Clustal Omega 11 1.2 Objectives 12 1.3 Scope of work 12 PART II MATERIALS AND METHODS 15 2.1 Equipment and Materials 15 2.1.1 Types of equipment 15 2.1.2 Materials 15 2.2 Methods 16 2.2.1 Growing cell A platensis C1 using Zarrouk’s medium [62] 16 2.2.2 Harvesting cell A platensis C1 by membranes filtration 17 2.2.3 Protein extraction from A platensis C1 18 2.2.4 Protein precipitation from A platensis C1 by acetone precipitation 19 2.2.5 Quantitative protein of A platensis C1 by 2-D Quant kit 20 2.2.6 Protein separation from A platensis C1 by Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS-PAGE) 21 2.2.7 In silico digestion tools 22 iii 2.2.8 ACE inhibition assay at Institute of Rangsit University 23 2.2.9 Common characteristics of antihypertensive peptides using Clustal Omega 24 PART III RESULTS AND DISCUSSIONS 25 3.1 Culture and cell growth of A platensis C1 25 3.2 Cell harvesting of A platensis C1 26 3.3 Protein extraction of A platensis C1 27 3.4 Protein precipitation from crude extract solution of A platensis C1 27 3.5 Protein quantification of A platensis C1 by 2D-Quant kit 28 3.6 Protein separation from the crude extract of A platensis C1 29 3.7 Common characteristics of antihypertensive peptides from A platensis C1 30 3.8 Discussion 36 PART IV CONCLUSION AND RECOMMENDATIONS 38 4.1 Conclusion 38 4.2 Recommendations 38 iv LIST OF TABLE Table 1.1 Amino acid profile of Arthrospira (Spirulina) platensis [60] Table 1.2 Genome statistics of A platensis C1 Table 1.3 Examples of bioactive peptides from Arthrospira (Spirulina) [38] Table 2.1 The equipment using for study 15 Table 2.2 The materials using for study 15 Table 2.3 Constituents of Zarrouk’s medium 16 Table 2.4 preparation of the standard curve 20 Table 3.1 The OD480 value of standard protein (BSA)and protein 28 of A platensis C1 28 Table 3.2 Classification of common amino acid of candidate antihypertensive peptides, discovered from A platensis C1 31 Table 3.3 Characteristics of Novel AHP based on ExactAHP_284 and ExactAHP_460 sequences 32 Table 3.4 Amount and percentage of functional amino acid groups of antihypertensive peptides from A platensis C1 34 Table 3.5 Number of amino acids found in Exactly_AHP of A platensis C1 35 Table 3.6 Functional amino acid pattern of Exactly_AHP of A platensis C1 35 v LIST OF FIGURES Fig 1.1 Classification of A platensisusing a cladistic approach [63] Fig 1.2 Morphological aspects of the evident cross-wall in Arthrospira isolated from soda Lake Kailala (Chad) (a) Fresh clonal trichome growth in the laboratory; (b) wild trichome fixed in formaldehyde (2%); (c) fresh clonal trichome grown in mass culture; (d) autofluorescence of trichome shown in (c); Bar maker = 10µm [7] Fig 1.3 Clonal trichomes of Arthrospira indicate isolated from Lake Lonar Arrows indicate the presence of a calyptra Bar marker = 20 mm [7] Fig 1.4 Viewed under a microscope, cells within A platensis trichomes are short and wide and have visible cross-walls [63] Fig 1.5 Life cycle of A platensis [15 Fig 1.6 Antihypertension mechanisms of marine algae-derived bioactive peptides that ACE and renin inhibitions are involved [22] Fig 1.7 Screenshot of the Clustal Omega Web page on the EBI Web site: 12 Fig 1.8 Schematic flow work in this report (blue boxes ); yellow boxes, done by Anekthanakul [61] 14 Fig 2.1 Process of cell A platensis culture in Zarrouk’s medium 16 Fig 2.2 Process of harvesting cell A platensis C1 by membrane filtration 17 Fig 2.3 Process of protein extraction from A platensis C1 18 Fig 2.4 Process of protein precipitation from A platensis C1 19 Fig 2.5 Process of quantitative protein of A platensis C1 21 Fig 2.6 Process of the separation protein of A platensis C1 22 Fig 2.7 Process of the in silico digestion and potential 22 Fig 2.8 Process of Antihypertension bioactivity testing 24 Fig 2.9 The Process of discovery for common characteristics of Novel AH peptide of A platensis C1 by Clustal Omega program 24 Fig 3.1 A platensis C1 grown culture in Zarrouk’s medium at35oC, 120 rpm under light illumination of 100 µE/m2/s Grown cells (day, OD560): (A) day 0, 0,1; (B) day 3, 0,742; (C) day 5, 1,463; and (D) day 7, 1,307 25 vi Fig 3.2 Growth curve of A platensis C1 cultured in Zarrouk’s medium at 35oC, 120 rpm under light illumination of 100 µE/m2/s 26 Fig 3.3 Harvesting of A platensis C1 cells by membranes filtration, (A) preparation of membranes on the filtration set, (B) adding about 250ml cell of A platensis C1, (C) harvested cells of A platensis C1 on the membrane before keeping them at -30oC 27 Fig 3.4 Protein extraction of A platensis C1 cells, (A) after sonicating, (B) after added dissolve buffer 27 Fig 3.5 Precipitation of crude protein extracted from A platensis C1: (A) precipitated protein after first centrifugation, (B) after precipitation eight times 28 Fig 3.6 Standard curve of protein standard (BSA) determined by the 2D-Quant kit method 29 Fig 3.7 Protein separation of samples (1 & 2) from A platensis C1 analyzed by 12% SDS-PAGE; (M), Markers 30 Fig 3.8 Common amino acid sequence of candidate antihypertensive peptideswith Novel AHP 33 Fig 3.9 Percentage of functional groups in the amino acid antihypertensive peptide sequences of A platensis C1 35 vii LIST OF ABBREVIATION ACE :Angiotensin Converting Enzyme HA : Hippuric acid AHP : Antihypertensive peptides BCG-cell : Bacillus Calmette–Guérin-cell BSA : Bovine Serum Albumin DS : Dissolve EPO :Erythropoietin ERK : Extracellular-signal Regulated Kinase HBP :High Blood Pressure HHL : Hippuryl-L-histidyl-L-leucine HPLC : High-performance liquid chromatography IC50 :Half maximal Inhibitory Concentration iNOS : Inhibits Nitric Oxide Synthase kD : Kilodalton LPS :Lipopolysaccharide MAPK : Mitogen-degradation Protein Kinase MSAs : Multiple Sequence Alignments M : Markers NF-kB : Nuclear Factor-kB OD :Optical Density PHB :Poly-β-Hydroxybutyrate RAS :Renin–Angiotensin System SDS-PAGE : Sodium Dodecyl Sulphate – Polyacrylamide Gel Electrophoresis WHO :World Health Organization viii ExactAHP_344 ExactAHP_171 ExactAHP_84 ExactAHP_320 ExactAHP_380 ExactAHP_374 ExactAHP_161 ExactAHP_106 ExactAHP_197 ExactAHP_396 ExactAHP_393 ExactAHP_389 ExactAHP_89 ExactAHP_388 ExactAHP_397 ExactAHP_372 ExactAHP_343 ExactAHP_307 ExactAHP_306 ExactAHP_169 ExactAHP_166 ExactAHP_168 ExactAHP_308 ExactAHP_309 ExactAHP_298 ExactAHP_300 ExactAHP_404 ExactAHP_295 ExactAHP_370 ExactAHP_82 ExactAHP_83 ExactAHP_297 ExactAHP_301 ExactAHP_412 ExactAHP_141 ExactAHP_342 ExactAHP_115 ExactAHP_114 ExactAHP_110 ExactAHP_113 ExactAHP_329 ExactAHP_311 ExactAHP_310 ExactAHP_153 ExactAHP_96 ExactAHP_59 ExactAHP_60 ExactAHP_416 ExactAHP_387 ExactAHP_352 ExactAHP_291 ExactAHP_279 ExactAHP_290 ExactAHP_314 ExactAHP_353 ExactAHP_457 ExactAHP_456 ExactAHP_400 ExactAHP_359 ExactAHP_355 ExactAHP_333 ExactAHP_266 ExactAHP_179 ExactAHP_406 ExactAHP_438 ExactAHP_439 ExactAHP_440 ExactAHP_434 ExactAHP_278 ExactAHP_229 ExactAHP_204 ExactAHP_163 ExactAHP_203 ExactAHP_283 ExactAHP_284 Novel AHP ExactAHP_460 ExactAHP_452 ExactAHP_449 ExactAHP_194 ExactAHP_92 ExactAHP_74 ExactAHP_91 ExactAHP_421 ExactAHP_422 ExactAHP_395 ExactAHP_394 ExactAHP_338 ExactAHP_253 SQPK -IQP FQP -QP VKP -VIKP IKP GKP -KP VSP VRSP -VPK -FVP -VP VVPP VHIPP SPP -PPK PP IPP IP -IPF -PQ -PQR PG -PGR VWP PER VFPS FP -FPE -PFPE -PH -WHP -HP SHP -GPV GPR GP -GPM RPR -PRVF -PR -IEP GEP -EP -EPR -WNPR -VNP TNP -NPP -MNP -NP PT TP -YVE YTVF VVY TVY -TQVY RVY LVY IVY -VY -YN -YNK YNKL YKY MKY LKY -KYY -IKY -KY -MY -MYY IRDLDYY -YY -YQY YQ -KFY -FYQQ FFYY FY -YA -YAAAT VRYL -VRY -RYQKF -LRY - ExactAHP_336 ExactAHP_437 ExactAHP_435 ExactAHP_433 ExactAHP_431 ExactAHP_376 ExactAHP_326 ExactAHP_223 ExactAHP_184 ExactAHP_22 ExactAHP_182 ExactAHP_183 ExactAHP_172 ExactAHP_66 ExactAHP_155 ExactAHP_32 ExactAHP_18 ExactAHP_346 ExactAHP_419 ExactAHP_420 ExactAHP_454 ExactAHP_405 ExactAHP_365 ExactAHP_39 ExactAHP_211 ExactAHP_143 ExactAHP_269 ExactAHP_293 ExactAHP_429 ExactAHP_430 ExactAHP_459 ExactAHP_424 ExactAHP_425 ExactAHP_426 ExactAHP_428 ExactAHP_427 ExactAHP_107 ExactAHP_133 ExactAHP_217 ExactAHP_104 ExactAHP_218 ExactAHP_402 ExactAHP_337 ExactAHP_159 ExactAHP_130 ExactAHP_49 ExactAHP_129 ExactAHP_195 ExactAHP_277 ExactAHP_78 ExactAHP_132 ExactAHP_103 ExactAHP_101 ExactAHP_102 ExactAHP_56 ExactAHP_371 ExactAHP_126 ExactAHP_125 ExactAHP_123 ExactAHP_124 ExactAHP_160 ExactAHP_157 ExactAHP_105 ExactAHP_100 ExactAHP_99 ExactAHP_77 ExactAHP_76 ExactAHP_47 ExactAHP_75 ExactAHP_318 ExactAHP_450 ExactAHP_351 ExactAHP_122 ExactAHP_121 ExactAHP_69 ExactAHP_120 ExactAHP_97 ExactAHP_98 ExactAHP_48 ExactAHP_339 ExactAHP_455 ExactAHP_423 ExactAHP_378 ExactAHP_362 ExactAHP_119 ExactAHP_10 ExactAHP_13 ExactAHP_15 ExactAHP_93 -RY YLLF -YL -YKW YK -VIY RIY LIY -IYLL AIYK IY -IYK -IQY -EY IEY ASY -AHSY -SY -WYS WYSL YS -VWY VAY -AY LAY -HY -LY -NY YH -YI -YW YG -YGG YGGY -YGLF -YGL -GL HGLF -LG GI LGI VWIG -RYLGY IG GYK DIGYY GY -KG MKG -FHG -HG GHG GH -GHF EG VG -GVW GVHGV -GV -GVGY -IGGSI -IFG GK -GGY GG FGK FGG -DFG -FG -QG YQGS TG GTW GTQY FATGT -GT -GF -GFHI DG SDGS YTAGV -YAGGS -VKAGF -VAGTW GS AG -AGS AGSS GA - ExactAHP_94 ExactAHP_95 ExactAHP_109 ExactAHP_116 ExactAHP_117 ExactAHP_127 ExactAHP_276 ExactAHP_286 ExactAHP_325 ExactAHP_341 ExactAHP_410 ExactAHP_177 ExactAHP_356 ExactAHP_357 ExactAHP_349 ExactAHP_354 ExactAHP_288 ExactAHP_199 ExactAHP_409 ExactAHP_58 ExactAHP_335 ExactAHP_23 ExactAHP_148 ExactAHP_144 ExactAHP_145 ExactAHP_411 ExactAHP_180 ExactAHP_181 ExactAHP_37 ExactAHP_271 ExactAHP_281 ExactAHP_334 ExactAHP_54 ExactAHP_282 ExactAHP_415 ExactAHP_417 ExactAHP_152 ExactAHP_65 ExactAHP_154 ExactAHP_162 ExactAHP_201 ExactAHP_142 ExactAHP_319 ExactAHP_135 ExactAHP_322 ExactAHP_174 ExactAHP_136 ExactAHP_173 ExactAHP_332 ExactAHP_331 ExactAHP_273 ExactAHP_198 ExactAHP_190 ExactAHP_57 ExactAHP_191 ExactAHP_188 ExactAHP_187 ExactAHP_50 ExactAHP_71 -GD -GE GM GQ -GR GW MG NG -RG SG -WG ITT -TTI TTN TE -TQ NK -KR -WA -EK RWAEK AKK -IAK IA -IAE WH IW -IWH AW MAW MRW -RW -DW -MW WM -WT -IE EW IEW IKW -KW -HQK -QK HIK RA IRA -HIR -IR -RS -RR ME -KQMK -KE EI KEIW -KAFR -KA DK FDKA - ExactAHP_165 ExactAHP_138 ExactAHP_61 ExactAHP_53 ExactAHP_30 ExactAHP_193 ExactAHP_1 ExactAHP_16 ExactAHP_19 ExactAHP_28 ExactAHP_45 ExactAHP_55 ExactAHP_340 ExactAHP_345 ExactAHP_323 ExactAHP_324 ExactAHP_156 ExactAHP_373 ExactAHP_176 ExactAHP_350 ExactAHP_81 ExactAHP_86 ExactAHP_44 ExactAHP_70 ExactAHP_46 ExactAHP_366 ExactAHP_369 ExactAHP_321 ExactAHP_178 ExactAHP_368 ExactAHP_72 ExactAHP_85 ExactAHP_131 ExactAHP_192 ExactAHP_274 ExactAHP_285 ExactAHP_418 ExactAHP_401 ExactAHP_403 ExactAHP_377 ExactAHP_379 ExactAHP_391 ExactAHP_292 ExactAHP_390 ExactAHP_367 ExactAHP_90 ExactAHP_87 ExactAHP_63 ExactAHP_364 ExactAHP_151 ExactAHP_36 ExactAHP_33 ExactAHP_34 ExactAHP_360 ExactAHP_361 ExactAHP_317 ExactAHP_6 ExactAHP_62 -INSQ -HK ER -DM -AR -KFAR -AA -AH -AI -AMQS -DA -EA SF -SWSF -RF -RFH IF -VIIF ITF -TF FNQ FSQ -CF FCF -DF VDF -VFK QVVF IVF -VF FE -FR HF -KF -MF -NF WV VW -VWIS -VK -VKK VRK -NVR -VR -VE FWN FTTQ EV VAV IAV -AVV AV -AVF -VAA VAF QAFT AF -ERAF - APPENDIX C Result amount and percentage of functional amino acid groups of antihypertensive peptides from A platensisC1 Aliphatic Aromatic Acidic Basic Hydroxylic Sulphur-containing Amidic Sequence name ExactAHP_1 ExactAHP_2 ExactAHP_3 ExactAHP_4 ExactAHP_5 ExactAHP_6 ExactAHP_7 ExactAHP_8 ExactAHP_9 ExactAHP_10 ExactAHP_11 ExactAHP_12 ExactAHP_13 ExactAHP_14 ExactAHP_15 ExactAHP_16 ExactAHP_17 ExactAHP_18 ExactAHP_19 Peptide sequence AA AAATP AAP ACHP AEL AF AFL AFLL AFP AG AGLP AGP AGS AGSP AGSS AH AHHP AHSY AI 675 314 56 168 70 27 50 49,63% 23,09% 4,12% 12,35% 5,15% 1,99% 3,68% Group A G I L P V F W Y D E R H K S T C M N Q A P P P L A L L P P P P G P G A P Y A 1 1 1 1 1 1 1 1 0 0 0 0 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 1 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Group of AH Aliphatic Aliphatic and Hydroxylic Aliphatic Aliphatic and Basic and Sulphur-containing Aliphatic and Acidic Aliphatic and Aromatic Aliphatic and Aromatic Aliphatic and Aromatic Aliphatic and Aromatic Aliphatic Aliphatic Aliphatic Aliphatic and Hydroxylic Aliphatic and Hydroxylic Aliphatic and Hydroxylic Aliphatic and Basic Aliphatic and Basic Aliphatic and Aromatic and Basic and Hydroxylic Aliphatic ExactAHP_20 ExactAHP_21 ExactAHP_22 ExactAHP_23 ExactAHP_24 ExactAHP_25 ExactAHP_26 ExactAHP_27 Peptide sequence AIP AIPP AIYK AKK ALEP ALPHA ALPM ALPP ExactAHP_28 AMQS A 0 0 0 0 0 0 0 1 ExactAHP_29 ExactAHP_30 ExactAHP_31 ExactAHP_32 ExactAHP_33 ExactAHP_34 ExactAHP_35 ExactAHP_36 ExactAHP_37 ExactAHP_38 ExactAHP_39 ExactAHP_40 ExactAHP_41 ExactAHP_42 ExactAHP_43 ExactAHP_44 ExactAHP_45 ExactAHP_46 ExactAHP_47 ExactAHP_48 ExactAHP_49 ExactAHP_50 ExactAHP_51 ExactAHP_52 ExactAHP_53 ExactAHP_54 ExactAHP_55 ExactAHP_56 AP AR ASP ASY AV AVF AVP AVV AW AWPF AY AYFYP AYP AYPQQ AYPS CF DA DF DFG DG DIGYY DK DL DLP DM DW EA EG P A P Y A A P A W P Y P P P P F A F G G G OR L P OR W A G 1 1 1 1 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 1 0 0 1 1 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Sequence name Group A G I L P V F W Y D E R H K S T C M N Q Group of AH P P Y A P P P P 1 1 1 0 0 0 0 1 0 0 0 0 1 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Aliphatic Aliphatic Aliphatic and Aromatic and Basic Aliphatic and Basic Aliphatic and Acidic Aliphatic and Basic Aliphatic and Sulphur-containing Aliphatic Aliphatic and Hydroxylic and Sulphur-containing and Amidic Aliphatic Aliphatic and Basic Aliphatic and Hydroxylic Aliphatic and Aromatic and Hydroxylic Aliphatic Aliphatic and Aromatic Aliphatic Aliphatic Aliphatic and Aromatic Aliphatic and Aromatic Aliphatic and Aromatic Aliphatic and Aromatic Aliphatic and Aromatic Aliphatic and Aromatic and Amidic Aliphatic and Aromatic and Hydroxylic Aromatic and Sulphur-containing Aliphatic and Acidic Aromatic and Acidic Aliphatic and Aromatic and Acidic Aliphatic and Acidic Aliphatic and Aromatic and Acidic Acidic and Basic Aliphatic and Acidic Aliphatic and Acidic Acidic and Sulphur-containing Aromatic and Acidic Aliphatic and Acidic Aliphatic and Acidic Sequence name ExactAHP_57 ExactAHP_58 ExactAHP_59 ExactAHP_60 ExactAHP_61 ExactAHP_62 ExactAHP_63 ExactAHP_64 ExactAHP_65 ExactAHP_66 ExactAHP_67 ExactAHP_68 ExactAHP_69 ExactAHP_70 ExactAHP_71 ExactAHP_72 ExactAHP_73 ExactAHP_74 ExactAHP_75 ExactAHP_76 ExactAHP_77 ExactAHP_78 ExactAHP_79 ExactAHP_80 ExactAHP_81 ExactAHP_82 ExactAHP_83 ExactAHP_84 ExactAHP_85 ExactAHP_86 ExactAHP_87 ExactAHP_88 ExactAHP_89 ExactAHP_90 ExactAHP_91 ExactAHP_92 ExactAHP_93 ExactAHP_94 Peptide sequence EI EK EP EPR ER ERAF EV EVLP EW EY FAL FAP FATGT FCF FDKA FE FFL FFYY FG FGG FGK FHG FLP FLPP FNQ FP FPE FQP FR FSQ FTTQ FVAP FVP FWN FY FYQQ GA GD Group A G I L P V F W Y D E R H K S T C M N Q OR OR P P OR A V P W Y L P G F OR F L Y G G G G P P F P P P F F OR P P OR Y Y G G 0 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 1 0 1 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 1 1 2 1 1 1 1 1 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 Group of AH Aliphatic and Acidic Acidic and Basic Aliphatic and Acidic Aliphatic and Acidic and Basic Acidic and Basic Aliphatic and Aromatic and Acidic and Basic Aliphatic and Acidic Aliphatic and Acidic Aromatic and Acidic Aromatic and Acidic Aliphatic and Aromatic Aliphatic and Aromatic Aliphatic and Aromatic and Hydroxylic Aromatic and Sulphur-containing Aliphatic and Aromatic and Acidic and Basic Aromatic and Acidic Aliphatic and Aromatic Aromatic Aliphatic and Aromatic Aliphatic and Aromatic Aliphatic and Aromatic and Basic Aliphatic and Aromatic and Basic Aliphatic and Aromatic Aliphatic and Aromatic Aromatic and Amidic Aliphatic and Aromatic Aliphatic and Aromatic and Acidic Aliphatic and Aromatic and Amidic Aromatic and Basic Aromatic and Hydroxylic and Amidic Aromatic and Hydroxylic and Amidic Aliphatic and Aromatic Aliphatic and Aromatic Aromatic and Amidic Aromatic Aromatic and Amidic Aliphatic Aliphatic and Acidic Sequence name ExactAHP_95 ExactAHP_96 ExactAHP_97 ExactAHP_98 ExactAHP_99 ExactAHP_100 ExactAHP_101 ExactAHP_102 ExactAHP_103 ExactAHP_104 ExactAHP_105 ExactAHP_106 ExactAHP_107 ExactAHP_108 ExactAHP_109 ExactAHP_110 ExactAHP_111 ExactAHP_112 ExactAHP_113 ExactAHP_114 ExactAHP_115 ExactAHP_116 ExactAHP_117 ExactAHP_118 ExactAHP_119 ExactAHP_120 ExactAHP_121 ExactAHP_122 ExactAHP_123 ExactAHP_124 ExactAHP_125 ExactAHP_126 ExactAHP_127 ExactAHP_128 ExactAHP_129 ExactAHP_130 ExactAHP_131 ExactAHP_132 Peptide sequence GE GEP GF GFHI GG GGY GH GHF GHG GI GK GKP GL GLP GM GP GPA GPL GPM GPR GPV GQ GR GRP GS GT GTQY GTW GV GVGY GVHGV GVW GW GWAP GY GYK HF HG Group A G I L P V F W Y D E R H K S T C M N Q G G G G G G G G G G G P G P G P P P P P P G G P G G G G G G G G G P G G F G 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 2 1 1 1 1 1 1 1 1 1 1 1 2 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Group of AH Aliphatic and Acidic Aliphatic and Acidic Aliphatic and Aromatic Aliphatic and Aromatic and Basic Aliphatic Aliphatic and Aromatic Aliphatic and Basic Aliphatic and Aromatic and Basic Aliphatic and Basic Aliphatic Aliphatic and Basic Aliphatic and Basic Aliphatic Aliphatic Aliphatic and Sulphur-containing Aliphatic Aliphatic Aliphatic Aliphatic and Sulphur-containing Aliphatic and Basic Aliphatic Aliphatic and Amidic Aliphatic and Basic Aliphatic and Basic Aliphatic and Hydroxylic Aliphatic and Hydroxylic Aliphatic and Aromatic and Hydroxylic and Amidic Aliphatic and Aromatic and Hydroxylic Aliphatic Aliphatic and Aromatic Aliphatic and Basic Aliphatic and Aromatic Aliphatic and Aromatic Aliphatic and Aromatic Aliphatic and Aromatic Aliphatic and Aromatic and Basic Aromatic and Basic Aliphatic and Basic Sequence name ExactAHP_133 ExactAHP_134 ExactAHP_135 ExactAHP_136 ExactAHP_137 ExactAHP_138 ExactAHP_139 ExactAHP_140 ExactAHP_141 ExactAHP_142 ExactAHP_143 ExactAHP_144 ExactAHP_145 ExactAHP_146 ExactAHP_147 ExactAHP_148 ExactAHP_149 ExactAHP_150 ExactAHP_151 ExactAHP_152 ExactAHP_153 ExactAHP_154 ExactAHP_155 ExactAHP_156 ExactAHP_157 ExactAHP_158 ExactAHP_159 ExactAHP_160 ExactAHP_161 ExactAHP_162 ExactAHP_163 ExactAHP_164 ExactAHP_165 ExactAHP_166 ExactAHP_167 ExactAHP_168 ExactAHP_169 ExactAHP_170 Peptide sequence HGLF HHL HIK HIR HIRL HK HL HLL HP HQK HY IA IAE IAIP IAIPP IAK IAP IAPG IAV IE IEP IEW IEY IF IFG IFL IG IGGSI IKP IKW IKY ILP INSQ IP IPA IPF IPP IPY Group A G I L P V F W Y D E R H K S T C M N Q G L OR R L OR L L P OR Y A A P P A P P A OR P W Y F G L G G P W Y P OR P P P P P 0 0 0 0 0 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 1 0 0 0 1 2 1 1 1 1 1 1 1 1 1 1 1 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Group of AH Aliphatic and Aromatic and Basic Aliphatic and Basic Aliphatic and Basic Aliphatic and Basic Aliphatic and Basic Basic Aliphatic and Basic Aliphatic and Basic Aliphatic and Basic Basic and Amidic Aromatic and Basic Aliphatic Aliphatic and Acidic Aliphatic Aliphatic Aliphatic and Basic Aliphatic Aliphatic Aliphatic Aliphatic and Acidic Aliphatic and Acidic Aliphatic and Aromatic and Acidic Aliphatic and Aromatic and Acidic Aliphatic and Aromatic Aliphatic and Aromatic Aliphatic and Aromatic Aliphatic Aliphatic and Hydroxylic Aliphatic and Basic Aliphatic and Aromatic and Basic Aliphatic and Aromatic and Basic Aliphatic Aliphatic and Hydroxylic and Amidic Aliphatic Aliphatic Aliphatic and Aromatic Aliphatic Aliphatic and Aromatic Sequence name ExactAHP_171 ExactAHP_172 ExactAHP_173 ExactAHP_174 ExactAHP_175 ExactAHP_176 ExactAHP_177 ExactAHP_178 ExactAHP_179 ExactAHP_180 ExactAHP_181 ExactAHP_182 ExactAHP_183 ExactAHP_184 ExactAHP_185 ExactAHP_186 ExactAHP_187 ExactAHP_188 ExactAHP_189 ExactAHP_190 ExactAHP_191 ExactAHP_192 ExactAHP_193 ExactAHP_194 ExactAHP_195 ExactAHP_196 ExactAHP_197 ExactAHP_198 ExactAHP_199 ExactAHP_200 ExactAHP_201 ExactAHP_202 ExactAHP_203 ExactAHP_204 ExactAHP_205 ExactAHP_206 ExactAHP_207 ExactAHP_208 Peptide sequence IQP IQY IR IRA IRP ITF ITT IVF IVY IW IWH IY IYK IYLL IYP IYPR KA KAFR KAIP KE KEIW KF KFAR KFY KG KL KP KQMK KR KTAP KW KWL KY KYY LA LAA LALPP LAMA Group A G I L P V F W Y D E R H K S T C M N Q P Y R R P F T F Y W W Y Y Y P P OR OR P OR OR F A Y G L P OR OR P W L Y Y L L P L 0 0 0 0 0 0 0 1 0 0 0 0 0 0 2 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 1 1 1 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Group of AH Aliphatic and Amidic Aliphatic and Aromatic and Amidic Aliphatic and Basic Aliphatic and Basic Aliphatic and Basic Aliphatic and Aromatic and Hydroxylic Aliphatic and Hydroxylic Aliphatic and Aromatic Aliphatic and Aromatic Aliphatic and Aromatic Aliphatic and Aromatic and Basic Aliphatic and Aromatic Aliphatic and Aromatic and Basic Aliphatic and Aromatic Aliphatic and Aromatic Aliphatic and Aromatic and Basic Aliphatic and Basic Aliphatic and Aromatic and Basic Aliphatic and Basic Acidic and Basic Aliphatic and Aromatic and Acidic and Basic Aromatic and Basic Aliphatic and Aromatic and Basic Aromatic and Basic Aliphatic and Basic Aliphatic and Basic Aliphatic and Basic Basic and Sulphur-containing and Amidic Basic Aliphatic and Basic and Hydroxylic Aromatic and Basic Aliphatic and Aromatic and Basic Aromatic and Basic Aromatic and Basic Aliphatic Aliphatic Aliphatic Aliphatic and Sulphur-containing ExactAHP_209 ExactAHP_210 ExactAHP_211 ExactAHP_212 ExactAHP_213 ExactAHP_214 ExactAHP_215 ExactAHP_216 ExactAHP_217 ExactAHP_218 ExactAHP_219 ExactAHP_220 ExactAHP_221 ExactAHP_222 ExactAHP_223 ExactAHP_224 ExactAHP_225 ExactAHP_226 ExactAHP_227 Peptide sequence LAP LAQ LAY LAYFYP LEF LEPP LF LFRQ LG LGI LGP LHLP LIQP LIVTQ LIY LIYP LKA LKL LKP ExactAHP_228 LKPMN P 0 1 0 0 0 0 0 1 ExactAHP_229 ExactAHP_230 ExactAHP_231 ExactAHP_232 ExactAHP_233 ExactAHP_234 ExactAHP_235 ExactAHP_236 ExactAHP_237 ExactAHP_238 ExactAHP_239 ExactAHP_240 ExactAHP_241 ExactAHP_242 ExactAHP_243 ExactAHP_244 ExactAHP_245 LKY LLF LLNP LLP LLPSY LN LNF LNP LNPA LNQ LPF LPG LPHF LPN LPP LQ LQKW Y L P P P L L P P L P P P P P L L 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 2 2 1 1 1 1 1 1 0 1 0 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 0 0 0 0 1 Sequence name Group A G I L P V F W Y D E R H K S T C M N Q P L Y P L P L L G G P P P L Y P L L P 1 1 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 1 1 0 1 1 1 1 1 1 1 1 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 Group of AH Aliphatic Aliphatic and Amidic Aliphatic and Aromatic Aliphatic and Aromatic Aliphatic and Aromatic and Acidic Aliphatic and Acidic Aliphatic and Aromatic Aliphatic and Aromatic and Basic and Amidic Aliphatic Aliphatic Aliphatic Aliphatic and Basic Aliphatic and Amidic Aliphatic and Hydroxylic and Amidic Aliphatic and Aromatic Aliphatic and Aromatic Aliphatic and Basic Aliphatic and Basic Aliphatic and Basic Aliphatic and Basic and Sulphur-containing and Amidic Aliphatic and Aromatic and Basic Aliphatic and Aromatic Aliphatic and Amidic Aliphatic Aliphatic and Aromatic and Hydroxylic Aliphatic and Amidic Aliphatic and Aromatic and Amidic Aliphatic and Amidic Aliphatic and Amidic Aliphatic and Amidic Aliphatic and Aromatic Aliphatic Aliphatic and Aromatic and Basic Aliphatic and Amidic Aliphatic Aliphatic and Amidic Aliphatic and Aromatic and Basic and Amidic ExactAHP_246 ExactAHP_247 ExactAHP_248 ExactAHP_249 ExactAHP_250 ExactAHP_251 ExactAHP_252 ExactAHP_253 ExactAHP_254 ExactAHP_255 ExactAHP_256 ExactAHP_257 ExactAHP_258 ExactAHP_259 ExactAHP_260 ExactAHP_261 ExactAHP_262 ExactAHP_263 ExactAHP_264 ExactAHP_265 ExactAHP_266 ExactAHP_267 ExactAHP_268 ExactAHP_269 ExactAHP_270 ExactAHP_271 ExactAHP_272 ExactAHP_273 ExactAHP_274 Peptide sequence LQP LQQ LQQQ LQQQQ LQSW LRP LRW LRY LSA LSLP LSP LSPA LSPY LSW LTF LVE LVL LVP LVR LVRT LVY LVYP LW LY LYP MAW MDLA ME MF ExactAHP_275 MFDL L 0 0 0 0 0 0 0 ExactAHP_276 ExactAHP_277 ExactAHP_278 ExactAHP_279 ExactAHP_280 ExactAHP_281 ExactAHP_282 MG MKG MKY MNP MPF MRW MW G G Y P P W W 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 0 0 0 0 0 0 Sequence name Group A G I L P V F W Y D E R H K S T C M N Q P L L L L P L Y L P P P P L L L L P L L Y P L Y P W L OR F 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 1 1 1 1 1 1 1 0 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Group of AH Aliphatic and Amidic Aliphatic and Amidic Aliphatic and Amidic Aliphatic and Amidic Aliphatic and Aromatic and Hydroxylic and Amidic Aliphatic and Basic Aliphatic and Aromatic and Basic Aliphatic and Aromatic and Basic Aliphatic and Hydroxylic Aliphatic and Hydroxylic Aliphatic and Hydroxylic Aliphatic and Hydroxylic Aliphatic and Aromatic and Hydroxylic Aliphatic and Aromatic and Hydroxylic Aliphatic and Aromatic and Hydroxylic Aliphatic and Acidic Aliphatic Aliphatic Aliphatic and Basic Aliphatic and Basic and Hydroxylic Aliphatic and Aromatic Aliphatic and Aromatic Aliphatic and Aromatic Aliphatic and Aromatic Aliphatic and Aromatic Aliphatic and Aromatic and Sulphur-containing Aliphatic and Acidic and Sulphur-containing Acidic and Sulphur-containing Aromatic and Sulphur-containing Aliphatic and Aromatic and Acidic and Sulphurcontaining Aliphatic and Sulphur-containing Aliphatic and Basic and Sulphur-containing Aromatic and Basic and Sulphur-containing Aliphatic and Sulphur-containing and Amidic Aliphatic and Aromatic and Sulphur-containing Aromatic and Basic and Sulphur-containing Aromatic and Sulphur-containing Sequence name ExactAHP_283 ExactAHP_284 ExactAHP_285 ExactAHP_286 ExactAHP_287 ExactAHP_288 ExactAHP_289 ExactAHP_290 ExactAHP_291 ExactAHP_292 ExactAHP_293 ExactAHP_294 ExactAHP_295 ExactAHP_296 ExactAHP_297 ExactAHP_298 ExactAHP_299 ExactAHP_300 ExactAHP_301 ExactAHP_302 ExactAHP_303 ExactAHP_304 ExactAHP_305 ExactAHP_306 ExactAHP_307 ExactAHP_308 ExactAHP_309 ExactAHP_310 ExactAHP_311 ExactAHP_312 ExactAHP_313 ExactAHP_314 ExactAHP_315 ExactAHP_316 ExactAHP_317 ExactAHP_318 ExactAHP_319 ExactAHP_320 Peptide sequence MY MYY NF NG NILP NK NKL NP NPP NVR NY PAP PER PF PFPE PG PGL PGR PH PL PLG PLP PLW PP PPK PQ PQR PR PRVF PRY PSY PT PYK PYP QAFT QG QK QP Group A G I L P V F W Y D E R H K S T C M N Q Y Y F G P OR L P P V Y P P P P P P P P P P P P P P P P P P P P P P P A G OR P 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 2 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 1 1 Group of AH Aromatic and Sulphur-containing Aromatic and Sulphur-containing Aromatic and Amidic Aliphatic and Amidic Aliphatic and Amidic Basic and Amidic Aliphatic and Basic and Amidic Aliphatic and Amidic Aliphatic and Amidic Aliphatic and Basic and Amidic Aromatic and Amidic Aliphatic Aliphatic and Acidic and Basic Aliphatic and Aromatic Aliphatic and Aromatic and Acidic Aliphatic Aliphatic Aliphatic and Basic Aliphatic and Basic Aliphatic Aliphatic Aliphatic Aliphatic and Aromatic Aliphatic Aliphatic and Basic Aliphatic and Amidic Aliphatic and Basic and Amidic Aliphatic and Basic Aliphatic and Aromatic and Basic Aliphatic and Aromatic and Basic Aliphatic and Aromatic and Hydroxylic Aliphatic and Hydroxylic Aliphatic and Aromatic and Basic Aliphatic and Aromatic Aliphatic and Aromatic and Hydroxylic and Amidic Aliphatic and Amidic Basic and Amidic Aliphatic and Amidic Sequence name ExactAHP_321 ExactAHP_322 ExactAHP_323 ExactAHP_324 ExactAHP_325 ExactAHP_326 ExactAHP_327 ExactAHP_328 ExactAHP_329 ExactAHP_330 ExactAHP_331 ExactAHP_332 ExactAHP_333 ExactAHP_334 ExactAHP_335 ExactAHP_336 ExactAHP_337 ExactAHP_338 ExactAHP_339 ExactAHP_340 ExactAHP_341 ExactAHP_342 ExactAHP_343 ExactAHP_344 ExactAHP_345 ExactAHP_346 ExactAHP_347 ExactAHP_348 ExactAHP_349 ExactAHP_350 ExactAHP_351 ExactAHP_352 ExactAHP_353 ExactAHP_354 ExactAHP_355 ExactAHP_356 ExactAHP_357 ExactAHP_358 Peptide sequence QVVF RA RF RFH RG RIY RL RP RPR RPY RR RS RVY RW RWAEK RY RYLGY RYQKF SDGS SF SG SHP SPP SQPK SWSF SY TAP TAPY TE TF TG TNP TP TQ TQVY TTI TTN TVPY Group A G I L P V F W Y D E R H K S T C M N Q F R F F G Y L P P P R R Y W A Y G Y G F G P P P F Y P P T F G P P T Y T T P 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 2 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 2 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 Group of AH Aliphatic and Aromatic and Amidic Aliphatic and Basic Aromatic and Basic Aromatic and Basic Aliphatic and Basic Aliphatic and Aromatic and Basic Aliphatic and Basic Aliphatic and Basic Aliphatic and Basic Aliphatic and Aromatic and Basic Basic Basic and Hydroxylic Aliphatic and Aromatic and Basic Aromatic and Basic Aliphatic and Aromatic and Acidic and Basic Aromatic and Basic Aliphatic and Aromatic and Basic Aromatic and Basic and Amidic Aliphatic and Acidic and Hydroxylic Aromatic and Hydroxylic Aliphatic and Hydroxylic Aliphatic and Basic and Hydroxylic Aliphatic and Hydroxylic Aliphatic and Basic and Hydroxylic and Amidic Aromatic and Hydroxylic Aromatic and Hydroxylic Aliphatic and Hydroxylic Aliphatic and Aromatic and Hydroxylic Acidic and Hydroxylic Aromatic and Hydroxylic Aliphatic and Hydroxylic Aliphatic and Hydroxylic and Amidic Aliphatic and Hydroxylic Hydroxylic and Amidic Aliphatic and Aromatic and Hydroxylic and Amidic Aliphatic and Hydroxylic Hydroxylic and Amidic Aliphatic and Aromatic and Hydroxylic Sequence name ExactAHP_359 ExactAHP_360 ExactAHP_361 ExactAHP_362 ExactAHP_363 ExactAHP_364 ExactAHP_365 ExactAHP_366 ExactAHP_367 ExactAHP_368 ExactAHP_369 ExactAHP_370 ExactAHP_371 ExactAHP_372 ExactAHP_373 ExactAHP_374 ExactAHP_375 ExactAHP_376 ExactAHP_377 ExactAHP_378 ExactAHP_379 ExactAHP_380 ExactAHP_381 ExactAHP_382 ExactAHP_383 ExactAHP_384 ExactAHP_385 ExactAHP_386 ExactAHP_387 ExactAHP_388 ExactAHP_389 ExactAHP_390 ExactAHP_391 ExactAHP_392 ExactAHP_393 ExactAHP_394 ExactAHP_395 ExactAHP_396 Peptide sequence TVY VAA VAF VAGTW VAP VAV VAY VDF VE VF VFK VFPS VG VHIPP VIIF VIKP VIPEL VIY VK VKAGF VKK VKP VLIVP VLKP VLP VLPEI VLPIP VMP VNP VP VPK VR VRK VRP VRSP VRY VRYL VSP Group A G I L P V F W Y D E R H K S T C M N Q Y A A G P A Y F V F V P G P F P P Y V G V P P P P P P P P P P V V P P Y Y P 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 0 0 0 0 1 0 0 1 1 1 1 0 1 0 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Group of AH Aliphatic and Aromatic and Hydroxylic Aliphatic Aliphatic and Aromatic Aliphatic and Aromatic and Hydroxylic Aliphatic Aliphatic Aliphatic and Aromatic Aliphatic and Aromatic and Acidic Aliphatic and Acidic Aliphatic and Aromatic Aliphatic and Aromatic and Basic Aliphatic and Aromatic and Hydroxylic Aliphatic Aliphatic and Basic Aliphatic and Aromatic Aliphatic and Basic Aliphatic and Acidic Aliphatic and Aromatic Aliphatic and Basic Aliphatic and Aromatic and Basic Aliphatic and Basic Aliphatic and Basic Aliphatic Aliphatic and Basic Aliphatic Aliphatic and Acidic Aliphatic Aliphatic and Sulphur-containing Aliphatic and Amidic Aliphatic Aliphatic and Basic Aliphatic and Basic Aliphatic and Basic Aliphatic and Basic Aliphatic and Basic and Hydroxylic Aliphatic and Aromatic and Basic Aliphatic and Aromatic and Basic Aliphatic and Hydroxylic Sequence name ExactAHP_397 ExactAHP_398 ExactAHP_399 ExactAHP_400 ExactAHP_401 ExactAHP_402 ExactAHP_403 ExactAHP_404 ExactAHP_405 ExactAHP_406 ExactAHP_407 ExactAHP_408 ExactAHP_409 ExactAHP_410 ExactAHP_411 ExactAHP_412 ExactAHP_413 ExactAHP_414 ExactAHP_415 ExactAHP_416 ExactAHP_417 ExactAHP_418 ExactAHP_419 ExactAHP_420 ExactAHP_421 ExactAHP_422 ExactAHP_423 ExactAHP_424 ExactAHP_425 ExactAHP_426 ExactAHP_427 ExactAHP_428 ExactAHP_429 ExactAHP_430 ExactAHP_431 ExactAHP_432 ExactAHP_433 ExactAHP_434 Peptide sequence VVPP VVPPA VVRP VVY VW VWIG VWIS VWP VWY VY VYAP VYP WA WG WH WHP WL WLP WM WNPR WT WV WYS WYSL YA YAAAT YAGGS YG YGG YGGY YGL YGLF YH YI YK YKLRP YKW YKY Group A G I L P V F W Y D E R H K S T C M N Q P P P Y V G V P Y Y P P OR G W P L P W P W W Y Y Y Y G G G G G G Y Y Y P Y Y 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 2 1 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 1 0 0 2 0 0 0 1 0 1 0 0 0 0 0 0 0 0 2 2 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 1 1 1 1 1 1 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 1 1 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 Group of AH Aliphatic Aliphatic Aliphatic and Basic Aliphatic and Aromatic Aliphatic and Aromatic Aliphatic and Aromatic Aliphatic and Aromatic and Hydroxylic Aliphatic and Aromatic Aliphatic and Aromatic Aliphatic and Aromatic Aliphatic and Aromatic Aliphatic and Aromatic Aliphatic and Aromatic Aliphatic and Aromatic Aromatic and Basic Aliphatic and Aromatic and Basic Aliphatic and Aromatic Aliphatic and Aromatic Aromatic and Sulphur-containing Aliphatic and Aromatic and Basic and Amidic Aromatic and Hydroxylic Aliphatic and Aromatic Aromatic and Hydroxylic Aliphatic and Aromatic and Hydroxylic Aliphatic and Aromatic Aliphatic and Aromatic and Hydroxylic Aliphatic and Aromatic and Hydroxylic Aliphatic and Aromatic Aliphatic and Aromatic Aliphatic and Aromatic Aliphatic and Aromatic Aliphatic and Aromatic Aromatic and Basic Aliphatic and Aromatic Aromatic and Basic Aliphatic and Aromatic and Basic Aromatic and Basic Aromatic and Basic Sequence name ExactAHP_435 ExactAHP_436 ExactAHP_437 ExactAHP_438 ExactAHP_439 ExactAHP_440 ExactAHP_441 ExactAHP_442 ExactAHP_443 ExactAHP_444 ExactAHP_445 ExactAHP_446 ExactAHP_447 ExactAHP_448 ExactAHP_449 ExactAHP_450 ExactAHP_451 ExactAHP_452 ExactAHP_453 ExactAHP_454 ExactAHP_455 ExactAHP_456 ExactAHP_457 ExactAHP_458 ExactAHP_459 ExactAHP_460 Peptide sequence YL YLEPA YLLF YN YNK YNKL YP YPF YPI YPK YPLDL YPLDLF YPR YPRY YQ YQGS YQQP YQY YRPY YS YTAGV YTVF YVE YVP YW YY Group A G I L P V F W Y D E R H K S T C M N Q Y P Y Y Y Y P P P P P P P P Y Y P Y P Y G Y Y P Y Y 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 2 0 0 0 0 0 0 0 0 0 1 1 1 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 1 1 1 1 2 1 1 1 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 Group of AH Aliphatic and Aromatic Aliphatic and Aromatic and Acidic Aliphatic and Aromatic Aromatic and Amidic Aromatic and Basic and Amidic Aliphatic and Aromatic and Basic and Amidic Aliphatic and Aromatic Aliphatic and Aromatic Aliphatic and Aromatic Aliphatic and Aromatic and Basic Aliphatic and Aromatic and Acidic Aliphatic and Aromatic and Acidic Aliphatic and Aromatic and Basic Aliphatic and Aromatic and Basic Aromatic and Amidic Aliphatic and Aromatic and Hydroxylic and Amidic Aliphatic and Aromatic and Amidic Aromatic and Amidic Aliphatic and Aromatic and Basic Aromatic and Hydroxylic Aliphatic and Aromatic and Hydroxylic Aliphatic and Aromatic and Hydroxylic Aliphatic and Aromatic and Acidic Aliphatic and Aromatic Aromatic Aromatic ... UNIVERSITY UNIVERSITY OF AGRICULTURE AND FORESTRY LUONG TAI CHIEN Topic title: BIOACTIVE PEPTIDES DISCOVERY FROM SPIRULINA: CLASSIFICATION OF COMMON CHARACTERISTICS OF ANTIHYPERTENSIVE PEPTIDES BACHELOR... University of Agriculture and Forestry Major Biotechnology Student name Luong Tai Chien Student ID DTN1353150004 Bioactive peptides discovery from Spirulina: Classification Thesis title of common characteristics. .. Clustal Omega Grouping Commonamino acid group Common characteristics of both AH and Novel AHpeptide Fig 2.9 The Process of discovery for common characteristics of Novel AH peptide of A platensis C1