formulation stability and characterization of protein and peptide drugs

... Gevaert et al COFRADIC and protein modifications analysis of purine-binding proteins in a total lysate of human Jurkat T-cells [40] primary separation mAU 1400 COFRADIC analysis of protein processing ... features of ATP-binding proteins As expected, the majority of FSBA-labeled Jurkat proteins were known binders of small nucleotides, cofactors, or DNA and RNA molecules However, several proteins and ... isolate, for instance, phosphopeptides [24], N-glycosylated peptides 6278 [25], ubiquitinated peptides [26], and N-terminal peptides [27] COFRADIC as a peptide- sorting tool Our peptide- centric proteome...

Ngày tải lên: 18/02/2014, 16:20

... prokaryotic and eukaryotic signal peptides and prediction of their cleavage sites Protein Eng 10, 1–6 13 Song J, Quan H & Wang C (1997) Dependence of the anti-chaperone activity of protein disulphide ... integrated elution peak areas, and the PDI’s oxidase and isomerase activities were expressed as the initial rate of decrease of linear tx3a and the initial rate of increase of nTx3.1, respectively To ... of disulfide oxidase and isomerase based on the model of hirudin folding Anal Biochem 339, 94–103 Walker KW, Lyles MM & Gilbert HF (1996) Catalysis of oxidative protein folding by mutants of protein...

Ngày tải lên: 07/03/2014, 05:20

... Each of these phage displays a random 12-mer peptide on its coat, by virtue of its capsid pVIII protein being tethered to the peptide by a (Gly4Ser)3 peptide linker Each peptide is composed of ... fetal erythroid-specific partner protein [25] Because Fe65 is a ligand of the Alzheimer’s b-amyloid precursor protein (APP) [32,33] and the aberrations of CP2 and ⁄ or APP in neuronal cells promote ... and verification of the CP2-binding ability of REST and YY1 that contain the HXPR motif As stated above, none of the 12-mer peptide sequences exactly matched those of any known proteins Having...

Ngày tải lên: 16/03/2014, 18:20

... loss of the a-helix during aggregation Meanwhile, the broadening of this band and the increase of the band intensity at 1665 cm-1 implies the increase of the random-coil content in the protein ... and Q (Figures 10 and 11, Tables and 5) The bands near 1339 [31,32] and 758 [33] cm-1 have been found to be indicators of the hydrophobicity of the Trp environment, and a decrease in these band ... variation of the zeta potential of the BSA, Lys, and Mb systems versus the concentration of DMSO (A, A’, and A’’) and Q (B, B’, and B’’) With increasing DMSO concentration, the zeta potential values of...

Ngày tải lên: 11/08/2014, 00:23

... the FERM (band 4.1, ezrin, radixin, moesin) domain of PTPH1 is necessary for the inhibition of Mek, Erk, Jnk and AP-1 and also for localization of the phosphatase on the plasma membrane of Jurkat ... was recorded and mean values of left and right paws were used as reaction index for the individual animal A cut-off of 25 seconds was used to avoid tissue damage in case of absence of response ... activation and IL8 production [25,26] CARR injection in the hind paw of the mouse is one of the most commonly used models of inflammation and inflammatory pain and it has a biphasic profile [27]...

Ngày tải lên: 11/08/2014, 03:20

... repolymerization and therefore growth cone dynamics (Rashid et al., 2001) Another group of Cdk5 substrates are the intermediate and heavy chains of neurofilament proteins (NFM and NFH, respectively) NFM and ... Immunofluorescence microscopy 57 2.2.14 Miscellaneous techniques 58 IV 2.2.15 Statistical analysis and presentation of data 58 Results and Discussion 60 3.1 Isolation of p35-associated proteins and ... absence of the β subunit The functions of the β subunit are to confer stability, regulate the enzymatic activity of the holoenzyme and the specificity of the catalytic subunit (Faust and Montenarh,...

Ngày tải lên: 16/09/2015, 17:10

... Cu2+-bound and Zn2+-bound aggregates and between Ab(1–40) and Ab(1–42) aggregates, indicating that both the type of bound metal and the peptide sequence dictate the mode of peptide assembly Results and ... between 200 and 250 nm, and averaged over 10 scans with a bandwidth of nm, a response time of s, a pitch of 0.5 nm, and a scan rate of 20 nmÆs)1, in a mm quartz cuvette AFM A portion of each Ab ... either in the form of aggregates formed in the presence of Cu2+ [(i) and (iii)] or Zn2+ [(ii) and (iv)], or as fibrils formed in the presence of EDTA [(v) and (vi)] The amount of amyloid or amyloid-like...

Ngày tải lên: 30/03/2014, 01:20

... discovery and analysis parts of functional proteomics which is the handling of very complex protein mixtures A protein of interest will typically represent only a tiny fraction of the total protein ... approaches offer generic advantages for the analysis of protein complexes both in terms of dynamic range (i.e how little of one protein can be identified in the presence of how much of a mixture of other ... fusion protein has been widely used for the discovery and analysis of individual protein interactions and, to a lesser extent, protein complexes [20,21] In this approach, the protein of interest...

Ngày tải lên: 31/03/2014, 07:20

... (enzyme I of the PTS), the rst protein in the cascade of proteins forming the PTS, to HPr, the histidine-phosphocarrier protein HPr is the smallest protein in the protein cascade of the PTS and it ... vant Hoff analysis and those determined by uorescence and DSC (see before) Determination of the Th and Ts The values of the temperatures where the enthalpy and entropy are zero, Th and TsÂ, ... conformational stability of a protein In Protein Structure (Creighton, T.E., ed.), 2nd edn, pp 253259 Oxford University Press, Oxford 35 Privalov, P.L (1979) Stability of proteins: small globular proteins...

Ngày tải lên: 19/02/2014, 12:20

... PDI family proteins of yeast and animals [9–12], but this is the first report to confirm expression of such proteins in the ER of plants A CXXS motif and a CXXC motif in the N-terminal and C-terminal ... All of these proteins had thiol oxidoreductase activity in vitro and were ubiquitously expressed in the body of the plant GmPDIL-1, GmPDIM and GmPDIS-1 are unfolded protein response genes, and ... both luminal proteins Generally, PDI family proteins play important roles in folding and quality control of nascent polypeptides [25] In soybean cotyledon, large amounts of seed storage proteins...

Ngày tải lên: 18/02/2014, 11:20

... with GroEL from 15 and 30 lg of the cell lysate without (lanes and 3) or in the presence of ATP (lanes and 4) was detected with anti-UP12 Ig Samples (3 and lg of total proteins) of the cell extract ... Effect of addition of mM DNP on cell growth (h) and expression of UP12 (j) (B) Monitoring of cell growth (s) and amount of UP12 (d) before and after addition of 0.1 mM CCCP (C) Effect of temperature ... (s) and relative amount of UP12 (d) were measured for 12 samples of cells taken at the indicated time (B) Western blot analysis of proteins in samples collected in (A) lg and lg of total protein...

Ngày tải lên: 22/02/2014, 07:20

... mechanism of transport function and the structure of the carrier proteins For example, the rate limiting step of peptide transporters has still not yet been determined, and the number of transporters ... kinetic parameters and transport characteristics of Bip-[3H]Pro Results and Discussion Synthesis, chemical characterization and stability of Bip-Pro Figure shows the structure (Fig 1A) and the synthesis ... min)1Æmg of protein) 1 These parameters agree very well with those of previous reports [18] For Bip-Pro uptake, an apparent Kt of 7.6 ± 1.8 lm and a Vmax of 1.1 ± 0.1 nmolÆ30 min)1Æmg of protein) 1...

Ngày tải lên: 07/03/2014, 05:20

... GmPDIL-2 and proglycinin or b-conglycinin during the course of the folding process of these proteins Results Cloning and expression of GmPDIL-1 and GmPDIL-2 To clone the soybean ortholog of Arabidopsis ... region between b and b¢ in GmPDIL-1, and between b and b¢ and b¢ and a¢ in GmPDIL-2, the structure of these regions may be protease resistant The activity of recombinant GmPDIL-1 and GmPDIL-2, which ... [34] In the case of GmPDIL-2, 10 of the 11 introns had TTNAN, and the eighth intron possessed CTNAN Promoter regions of around kb and 1.5 kb upstream of each start codon of GmPDIL-1 and GmPDIL-2...

Ngày tải lên: 07/03/2014, 05:20

... 205 and 215 nm, indicating that Rv1399c is composed of 22% of a-helices, 25% of b-strand and 39% of random coil The catalytic triad is located at the bottom of a small ˚ ˚ pocket, 20 A length and ... sheet is composed of parallel b strands associated to an antiparallel strand (b2) and is surrounded by helices (a1, a2, a3, a7 and a8) The second domain consists of helices a4, a5 and a6 all clustered ... and Discussion Cloning and expression of Rv1399c The Rv1399c gene product encodes a 318 amino acid protein with a molecular mass of  31.7 kDa and a calculated pI of 4.53 and belongs to the Lip...

Ngày tải lên: 07/03/2014, 16:20

... Comparison of the fasciclin I domains in CLAP with selected proteins in the protein database The Conserved Domain Database of NCBI was screened with the protein coding sequence of clone of CLAP and ... Processing of the pro-CLAP C-terminus is essential to lower the molecular mass and pI of the protein to values observed by SDS/ PAGE and IEF Interestingly, pool sequencing of the mixture of CNBr peptides ... assigned to bands in columns e and f were determined from both IEF standards (Std) and buffer control gel run in parallel The numbers at the right represent the pI values of the major bands in e and f,...

Ngày tải lên: 07/03/2014, 16:20

... homologs of OsRPA70a and OsRPA70b (AtRPA70a and AtRPA70b) A thaliana was used for genetic analysis of the functions of OsRPA70a and OsRPA70b because it has closely related homologs (AtRPA70a and AtRPA70b, ... efficiency of the PCR (Fig 1B) We performed Western blot analysis of total proteins from seedlings of wild-type, atrpa70b, and the AtRPA70b RNAi mutant, and confirmed that very little AtRPA70b protein ... 20% of the relative activity of the OsRPA32 and OsRPA14 interaction The interaction of OsRPA70b and OsRPA32 yielded about 75% of the relative activity of the OsRPA32 and OsRPA14 interaction The...

Ngày tải lên: 07/03/2014, 21:20

... Van Hoof, C., Ingels, F., Cayla, X., Stevens, I., Merlevede, W & Goris, J (1995) Molecular cloning and developmental regulation of expression of two isoforms of the catalytic subunit of protein ... located However, this Xenopus protein has a long N-terminal extension, and so encodes a protein of about 70 kDa instead of 48 kDa Screening of Xenopus B¢¢ subunits of PP2A (Eur J Biochem 270) 385 ... a protein of about 70 kDa Since HPR72 also encodes a protein of about 70 kDa and PR70 and PR72 have quite similar amino acid sequences, they might fulfil overlapping functions Besides XPR130 and...

Ngày tải lên: 08/03/2014, 08:20

... (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4 Nature 227, 680 Ansorge, W (1985) Fast and sensitive detection of protein and DNA bands by treatment ... inhibition of translation) of lamjapin is about 0.69 nM, a very low value for an inhibitor of protein Table Purification procedures of lamjapin from 30 g dry powder of L japonica A One unit of specific ... pI values of the protein markers was used to calculate the pI value of lamjapin biosynthesis, and still in the range of the IC50 (0.03–4 nM) of type I RIPs The IC90 (the concentration of RIP causing...

Ngày tải lên: 08/03/2014, 10:20

... of calhepatin was aligned with those of the human form of each S100 protein, except for MRP126 and ictacalcin, which have only been isolated from chicken and catfish, respectively The number of ... dimerization and monomer stability, in Sl00 A4 and other S100 proteins [23] (Fig 3), are clustered between helix I and IV (Fig 5B), in the same way as in Sl00 A4 and other S100 proteins [23] ... C.W (1994) Purification and characterization of the recombinant human calcium-binding S100 proteins CAPL and CACY Biochemistry 33, 6732–6738 Moore, D.S (1985) Amino acid and peptide net charges:...

Ngày tải lên: 08/03/2014, 22:20

... PHM, GRF and GIP GLP-1 and GLP-2 are similar peptides, with molecular masses of 3299 Da and 3768 Da, respectively Both peptides have His-Ala as the N-terminal dipeptide, and DPP4 cleavage of these ... capable of the same cleavage event, producing peptides of 3090 Da and 3558 Da for GLP-1 and GLP-2, respectively Both peptides were inefficient FAP substrates, with half-lives of 22 h and 19 h, ... with peptide chain length [43] However, in contrast to this, in the case of GRF, the rate of DPP4 hydrolysis of longer peptides (44 amino acids) is higher than that of shorter peptides (three and...

Ngày tải lên: 14/03/2014, 23:20