imaging gpcr activation through monitoring dissociation of g protein heterotrimer in living cells
Báo cáo khoa học: C-terminal truncated cannabinoid receptor 1 coexpressed with G protein trimer in Sf9 cells exists in a precoupled state and shows constitutive activity ppt
Ngày tải lên :
23/03/2014, 07:20
... Precoupled form of human CB1 and G protein trimer G CB1(417) +G Anti-Flag M2 IgG Receptor Anti-His tag IgG Receptor Anti-His tag IgG G Anti-Gi1/Gi2 IgG G containing only G proteins did not show ... antagonist AM251 inhibited this GTP binding to G proteins The agonist WIN 55,212–2 increased GTP binding to a 6110 G Time (min) CB1 + G + IA CB1 + G + A CB1 + G 12 Fig Radioactive GTPcS binding assay ... and NotI restriction sites, was 5¢-GAA T GC GGC CGC TCA CTT TTC GAA TTG AGG GTG CGA CCA GAA TTC AGC CTC GGC AGA CGT GTC TGT GGA-3¢, which contains the StrepII tag between the EcoRI and NotI sites...
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optimization of protein and rna detection methodologies and a new approach for manipulating protein activity in living cells
Ngày tải lên :
13/11/2014, 10:46
... GGGCAGCACCACCGAGCGGATCCGGCCATGGT 31 ATGGCCGGATCCCATGCGTGGTGCTGCCCCGG 32 CCGGGGCAGCACCACGCATGGGATCCGGCCAT 33 GCCGGATCCCATCGGGCGTGCTGCCCCGGCTG 34 CAGCCGGGGCAGCACGCCCGATGGGATCCGGC 35 ACCATGGCCGGATCCGCTTTGAATTGCTGCCC 36 GGGCAGCAATTCAAAGCGGATCCGGCCATGGT ... 5’PHOS-GAACCGGAGCCGCAGCACCCGGGGCAGCAAGGCATT 27 GGGAAGCTTGCCACCATGGGTGACTGGAGTGCCTTGGGGAAATTACTGG ACAAGG 28 AAAAGGTACCGACCGGTTGAACCGCAATCTCCAGGTCATCAG 29 ACCATGGCCGGATCCGCTCGGTGGTGCTGCCC 30 GGGCAGCACCACCGAGCGGATCCGGCCATGGT ... GGGGCATATGCCAAGTACGCCCCCTATT 42 CGGCCGCCAAAAGCTGCGCAGCAGCCGGGGC 43 CTGCGGCCGCCAAAAGCCTTGCAGCAGCCGG 44 TTGCTGCGGCCGCCAGCAGTCTTGCAGCAGCC 45 CGGCCGCCCGGCTCCGCGCAGCAGCCGGGGCA 46 CTGCGGCCGCCCGGCGCCATGCAGCAGCCGG 47...
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Báo cáo khoa học: Activation of nematode G protein GOA-1 by the human muscarinic acetylcholine receptor M2 subtype Functional coupling of G-protein-coupled receptor and G protein originated from evolutionarily distant animals doc
Ngày tải lên :
07/03/2014, 11:20
... stage of C elegans as a template with the following primers: M2-goa1-s, 5¢-CATTATAAGA ACATAGGCGCTACAAGGATGGGTTGTACCATGTC ACAGGAAG-3¢; M2-goa1-PstI-as, 5¢-CCAATGCATTGG TTCTGCAGTTAATACAAGCCGCATCCACGAAGA-3¢ ... conditions Human GPCR activates nematode G protein construct, pPAK-M2–Gai1 [25], as a template with the following primers: M2-myc-EcoRI-s, 5¢-CAGAATTCatg gagcagaagctgatctccgaggaggacctgctgGTGAACAACTCCAC ... structure determination in 1986 [23] Although Gb and Gc are essential for Ga activation by GPCR [10], GPCR can activate Ga without Gb and Gc in some GPCR: :Ga fusion proteins [24] Muscarinic-agonist-dependent...
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Báo cáo y học: "Expression of a protein involved in bone resorption, Dkk1, is activated by HTLV-1 bZIP factor through its activation domain" potx
Ngày tải lên :
13/08/2014, 01:20
... DKK1-1853F, 5′TGGAATTTGGGATGGGAAGGACAC; DKK1-1854R, 5′-CACCACCAAGTAAAGCCAGTGACA; DKK1-991F, 5′-CATTCGGAAGCGTTGCGATGTGAT; DKK1-991R, 5′-ACTTGATTAGGCAGACGCGTGAGA; DKK1-331F, 5′-ACTTGTGTGCACAGTCAGCGAGTA; ... 5′-ATGGCCCACTTC CCAGGGTTTGGA; Tax-R, 5′-ACCAGTCGCCTTGTACACAGTCTC; HBZ-S1-F, 5′- TTAAACTTACCTAGACGGCGGACG; HBZ-S1-R, 5′-GCATGACACAGG CAAGCATCGAAA; ACTB-F, 5′-ACCAACTGGGACGACATGGAGAAA; ACTBR, 5′-TAGCACAGCCTGGATAGCAACGTA ... 5′-TTAATAAATGCAGGCGGCAGCAGG; DKK1 + 33F, 5′-AAATCCCATCCCGGCTTTGTTGTC; DKK1 + 33R, 5′-TCTCAGAAGGACTCAAGAGGGAGA Polakowski et al Retrovirology 2010, 7:61 http://www.retrovirology.com/content/7/1/61 Page of...
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CHARACTERIZATION OF g PROTEIN COUPLED RECEPTORS THROUGH THE USE OF BIO AND CHEMO INFORMATICS TOOLS
Ngày tải lên :
31/10/2015, 10:39
... ORs, greatly limiting our understanding of olfactory coding In 2009, Saito and coworkers, have performed high-throughput screening of 93 odorants against 464 ORs expressed in heterologous cells ... homology modeling and the built web-server automatically modeling of GPCRs at different activation states automatic modeling of GPCRs at different activation states, and docking odorant ligands II WEB-BASED ... of GPCRs through the use of bioand chemo- informatics tools The server will play the role for doing homology modeling massively for the unknown 3D GPCRs based on the experimentally known 3D GPCRs...
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Tài liệu Báo cáo khoa học: Regulators of G-protein signalling are modulated by bacterial lipopeptides and lipopolysaccharide pptx
Ngày tải lên :
18/02/2014, 13:20
... to give the inactive heterotrimeric form [15] Regulators of G- protein signalling (RGS) interact directly with the G protein a subunit in order to inhibit G- protein signalling [16] RGS proteins ... muRGS1 5¢-TCTGCTAGCCCAAAGGATTC-3¢ (sense), 5¢TTCACGTCCATTCCAAAAGTC-3¢ (anti-sense); muRGS2 5¢-GAGAAAATGAAGCGGACACTCT-3¢ (sense), 5¢-TTG CCAGTTTTGGGCTTC-3¢ (antisense); muHPRT as housekeeping gene ... 12 13 14 15 16 17 658 S Riekenberg et al of regulator of G protein signaling proteins in dendritic cells: implications for G protein- coupled receptor signaling J Immunol 172, 5175–5184 Hoebe...
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Báo cáo khoa học: Identification of the structural determinant responsible for the phosphorylation of G-protein activated potassium channel 1 by cAMP-dependent protein kinase pdf
Ngày tải lên :
07/03/2014, 00:20
... Autoradiograms of subsequent SDS gels revealed GIRK1 migrating in two bands, indicating a glycosylated and a nonglycosylated form, as reported previously [24] Interestingly, prominent in vitro ... mutations to abolish protein phosphorylation was a result of other protein kinases masking the PKA-catalysed part Indeed, protein kinases other than PKA, including 6222 tyrosine kinases, have been ... in (A), but with currents originating from the triple-mutated GIRK1F137S*** protein (C) Statistics of the effects of cAMP on basal currents of heterooligomeric GIRK1 ⁄ GIRK4, homooligomeric GIRK1...
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Báo cáo khoa học: The variable C-terminal extension of G-protein-coupled receptor kinase 6 constitutes an accessorial autoregulatory domain ppt
Ngày tải lên :
07/03/2014, 12:20
... single incubation at 72 °C for 10 min) using primer P1, 5¢-AGCCCATGGAGCTCGAGAACA TCGTA-3¢ (nucleotides 1–26, sense, initiating ATG underlined) in combination with antisense primers introducing ... binding of a C-terminal region including a pleckstrin homology domain to membrane-bound G protein bc dimers and negatively charged membrane phospholipids, including PtdInsP2 [30–34] Mutations in ... codon sequence: mGRK6-A M2: P2, 5¢-CTAGTCGC TGGAGTTCCCAGAGGAATCTTGGCG-3¢ (nucleotides 1677–1706, antisense, stop codon underlined) and mGRK6A M3: P3, 5¢-CTAATCTTGGCGACTGAAGAGTCT-3¢ (nucleotides...
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Báo cáo khoa học: Ternary complex formation of pVHL, elongin B and elongin C visualized in living cells by a fluorescence resonance energy transfer–fluorescence lifetime imaging microscopy technique docx
Ngày tải lên :
16/03/2014, 05:20
... 5¢-CTAGAC CACCATGTACCCCTACGACGTGCCCGACTACGCCG ATATCCCGGGTTAACT-3¢ and 5¢-CTAGAGTTAACC CGGGATATCGGCGTAGTCGGGCACGTCGTAGGGG TACATGGTGGT-3¢, into the XbaI site of pBOS Vector pBOS-Myc and pBOSFlag were constructed ... similarly by using the synthesized oligonucleotides 5¢-CTAGACCA CCATGGAGGAACAGAAGCTGATCAGTGAGGAAG ACCTGGATATCCCGGGTTAACT-3¢ and 5¢-CTAGAG TTAACCCGGGATATCCAGGTCTTCCTCACTGATCA GCTTCTGTTCCTCCATGGTGGT-3¢, ... 5¢-CGTGACCACCCTGACCGCGGG CGTGCAGTGCTTC-3¢ and 5¢-GAAGCACTGCACGCC CGCGGTCAGGGTGGTCACG-3¢ pCerulean(W66A)elongin C-citrine was constructed by inserting the BsrGI-EcoRI fragment of pcerulean-elongin C and...
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Báo cáo khóa học: Nerve growth factor mediates activation of the Smad pathway in PC12 cells doc
Ngày tải lên :
16/03/2014, 16:20
... SSXS-motif Involvement of Smad4 in NGF-triggered activation of the Smad signaling cascade In TGF-b1-induced signaling, R-Smads form heteromeric complexes with Smad4 following the activation by TbRI Fig ... transfection of PC12 cells with TbRII but not TbRI constructs leads to increased responsiveness to TGF-b1 (Fig 2B), again indicating that TbRII is the limiting component of TGF-b1 signaling in PC12 cells ... (1997) TGF-b signalling from cell membrane to nucleus through Smad proteins Nature 390, 465–471 ´ 12 Massague, J & Chen, Y .G (2000) Controlling TGF-b signaling Genes Dev 14, 627–644 ´ 13 Lagna, G. ,...
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Báo cáo khoa học: The impact of G-protein-coupled receptor hetero-oligomerization on function and pharmacology pptx
Ngày tải lên :
16/03/2014, 22:20
... hetero-oligomers A B H GPCR GPCR H GPCR H H β-arrestin H GPCR H H GPCR GPCR H GPCR GPCR β-arrestin No effect No effect H GPCR H GPCR H GPCR β-arrestin β-arrestin H GPCR β-arrestin ERK activation ERK activation ... cartoon showing the hypothetical mechanisms of receptor–b-arrestin interaction and ERK1 ⁄ signaling is shown in Fig Regardless of the mechanism by which b-arrestins bind to GPCRs, the signaling pathway ... effects on dampening receptor G- protein coupling and on receptor internalization, b-arrestin also plays a major role in GPCR activation of mitogen-activated protein kinase (MAPK) In this context...
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Báo cáo khoa học: The study of G-protein coupled receptor oligomerization with computational modeling and bioinformatics doc
Ngày tải lên :
16/03/2014, 22:20
... effects of dimerization ⁄ oligomerization on mechanisms of ligand binding and extent of activation (i.e pharmacological efficacy), as well as on the involvement of protein protein interactions in GPCR ... Domain swapping in G- protein coupled receptor dimers Prot Eng 11, 1181–1193 Gouldson PR, Higgs C, Smith RE, Dean MK, Gkoutos GV & Reynolds CA (2000) Dimerization and domain swapping in G- protein- coupled ... compensated by changes in the interacting protein (Fig 2C) to preserve the protein protein interface In the hypothetical multiple sequence alignment shown in Fig 2, these compensatory changes occur at...
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Báo cáo khoa học: Methods to monitor the quaternary structure of G protein-coupled receptors doc
Ngày tải lên :
16/03/2014, 22:20
... identification of potential protein protein interaction sites in GPCR dimers [22,23] and determining whether ligand-binding modulates the organizational structure of a GPCR dimer ⁄ oligomer Although the ... higher concentration of G protein provided in the membranes by the : GPCR : G protein stoichiometry of the fusion proteins allowing the G protein fused to one receptor to interact with the G protein ... ligand-binding, studies that have used pairs of nonfunctional mutants to restore GPCR signalling have produced data consistent with GPCR GPCR interactions By generating mutants of the luteinizing hormone...
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báo cáo khoa học: " Nucleoside conjugates of quantum dots for characterization of G protein-coupled receptors: strategies for immobilizing A2A adenosine receptor agonists" pdf
Ngày tải lên :
11/08/2014, 00:22
... anchoring moiety for chains containing a single nucleoside moiety The route in Figure utilized an exclusively amide-linked chain, and in Figure an intervening poly(ethyleneglycol) (PEG) spacer group ... 5'-N-ethylcarboxamidoadenosine (NECA) Each tube in the binding assay contained 100 μL of membrane suspension (20 g of protein) , 50 μL of agonist radioligand, and 50 μL of increasing concentrations of the test ligands ... coupling of 1a through a long-chain PEG spacer group of ten units present in Das et al Journal of Nanobiotechnology 2010, 8:11 http://www.jnanobiotechnology.com/content/8/1/11 Page of 19 Figure...
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Báo cáo y học: " Signaling and regulation of G protein-coupled receptors in airway smooth muscle" ppsx
Ngày tải lên :
13/08/2014, 13:20
... discovered RGS (regulators of G protein signaling) proteins [107] Experimental manipulation of RGS protein expression can alter GPCR signaling, but the physiologic role of RGS proteins is unclear Interestingly, ... (PKA) or protein kinase C (PKC) can also phosphorylate GPCRs and promote a loss of GPCR- G protein coupling See text for details regulation of GPCRs in ASM, the trafficking of GPCRs to their degradation ... analyzing GPCR signaling in ASM run the spectrum of integrative to reductionist approaches, each having certain advantages and disadvantages Integrative in vivo models in which GPCR ligands are...
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Near infrared autofluorescence imaging and spectroscopy for early detection of precancer and cancer in the colon
Ngày tải lên :
10/09/2015, 08:29
... during clinical colonoscopic screening V List of Figures Fig 1.1 Anatomy of the colon…………….……………………………… …… Fig 1.2 Conceptualization of morphologic progression through oncogenesis, incorporating ... detail by spraying of stains, thus it has been widely applied in a variety of clinical settings and throughout all gastrointestinal tract segments (including the colon) by the endoscope in the past ... cancer detection and diagnosis The ratio imaging of NIR DR image to NIR AF image is used to eliminate the geometric effect of the imaging measurement and the depolarization imaging is expected to improve...
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Analysis of regulator of g protein signaling (RGS) function in growth, development and pathogenicity of magnaporthe grisea 1
Ngày tải lên :
14/09/2015, 09:13
... subunit GAP Guanosine triphosphatase activating proteins GDP Guanosine diphosphate GEF Guanine-nucleotide exchange factors GIRK G protein- coupled inwardly rectifying potassium G protein Guanine nucleotide ... regulator of G protein signaling (RGS), leads to the replacement of GTP with GDP and reassociation of G with G γ heterodimer, leading to the termination of G protein- mediated signaling (Figure 3) ... Desensitization of G protein Signaling 1.2.3.1 The discovery of Regulator of G protein signaling (RGS) Studies of the desensitization of G- protein- coupled signal transduction have led to the discovery of...
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Analysis of regulator of g protein signaling (RGS) function in growth, development and pathogenicity of magnaporthe grisea 2
Ngày tải lên :
14/09/2015, 09:24
... 466 QQDRSHIQQFPGCQVFQPTKHAIYHMTSKGKDMINGSVPRGRASEGDATHSATHRHG-VA QQDRAYTAQYPGSQLFQPTKHSIYQITPNGKDLINGMNSRGRTSDAESTPRDHKPNEKLP QEDKGYPQPDASIVVFQPSKYAIYGITERGQRVCG -WIARDKSRETFYDNRGMP LIDSKHCDKKSNTSTSKNNIVKTIDSALMKQANECLEMAYHIYSSYIMIGSPYQLNIHHN ... HPH1 RGS1 Figure 21 A rgs1D WT Rgs1 OP Complemented B WT rgs1D Complemented Rgs1 OP _ Figure 22 Inductive Non-inductive _ Figure 23 WT rgs1D Complemented Figure 24 Figure 25 Figure 26 Rgs1 MG00990 ... ESPRNTMQLVNLERDTETDKLSHDRATIEVIFRRFAGQDGPNVKSSISTSDSDSLSDYSN FQISRSSFFTLSKRGWDLVSWTGCKSNNIRAPNGSTIDLDFTLRGHMTVRDEKKTLDDSE Rgs1 Cprgs-1 FlbA Sst2 408 213 420 406 GLTGVKMAPERKVNGKIHKDTFTGK-AASEWLMDCCTTVDRREAVEIASLFVEYELIEAL GLTGVKMAAERKIGGKTYKETFTGK-AATDWLMDCSTTVDRRETVEIASYFVEFGLMECV...
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Analysis of regulator of g protein signaling (RGS) function in growth, development and pathogenicity of magnaporthe grisea 3
Ngày tải lên :
14/09/2015, 09:39
... Figure 28 Inductive Non-inductive _ Figure 29 Figure 30 Inductive Non-inductive _ Figure 31 B A Figure 32 A B Figure 33 A B Figure 34 mgb1D WT _ rgs1Dmgb1D magB G1 83Smgb1D Figure 35 Figure ... mgb1D WT _ rgs1Dmgb1D magB G1 83Smgb1D Figure 35 Figure 36 MG010315 MG010105 MG09134 Control: Gamma actin MG03982 MG01173 MG01630 Figure 37 A B ...
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Analysis of regulator of g protein signaling (RGS) function in growth, development and pathogenicity of magnaporthe grisea 4
Ngày tải lên :
14/09/2015, 09:53
... Figure 39 WT rgs1D Figure 40 Figure 41 Figure 42 _ Figure 43 - Gd + Gd, 1h + Gd, 0h + Gd, 2h + Gd, 0.5h + Gd, 3h _ Figure 44 A WT rgs1D Solvent Solvent 10mM 8-Br-cAMP 10mM 8-Br-cAMP _ B WT rgs1D ... Solvent 10mM 8-Br-cAMP 10mM 8-Br-cAMP _ B WT rgs1D Figure 45 Appressorium formation on inductive membrane (%) Appressorium formation on Noninductive membrane (%) WT 89 1.5 mscL∆ 92 mscS1∆ 87 2.5...
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