involving g protein coupled receptors gpcr

... Class C G- protein- coupled receptors ligand binding This led to the demonstration that most GPCRs can oligomerize as shown by both biochemical and energy transfer technologies [3] In recent ... intracellular loops of class C GPCRs as well as the C-terminal tail are involved in G- protein coupling For various class C GPCRs, including the mGlu5, GABAB2 and CaS receptors, the HD can fold correctly ... occurs in the GABAB receptor, GABA binding in the GABAB1 VFT leading to activation of the GABAB2 HD Although GABAB1 VFT binds the agonist and the GABAB2 HD couples to G- protein, a chimeric construct...

Ngày tải lên: 07/03/2014, 21:20

... very high GPCR- GFP ⁄ GPCR- Rluc ratios (B) BRET2 measurements performed in cells expressing increasing concentrations of GPCR Rluc and GPCR GFP2 while maintaining a constant GPCR GFP2 : GPCR Rluc ... single GPCR, Carrillo et al [61] extended the use of GPCR -G protein fusion proteins [62] to study both homo- and hetero dimerization of GPCRs Mutations were introduced to generate pairs 2921 GPCR ... restoration of 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...

Ngày tải lên: 16/03/2014, 22:20

... KA: Functionalized congener approach to the design of ligands for G protein- coupled receptors (GPCRs) Bioconjugate Chem 2009, 20:1816-1835 10 Kim Y, Hechler B, Klutz AM, Gachet C, Jacobson KA: ... linker (Figure 8) was even greater than that of the dendron-nucleoside conjugate 11, suggesting that loss of affinity is not a necessary consequence upon tethering a small molecular GPCR ligand to ... Toward multivalent signaling across G protein- coupled receptors from poly(amidoamine) dendrimers Bioconjugate Chem 2008, 19:406-411 11 Klutz KM, Gao ZG, Lloyd J, Shainberg A, Jacobson KA: Enhanced...

Ngày tải lên: 11/08/2014, 00:22

... and actin polymerization [40] strongly suggest a physiologic role for G1 2/13 signaling in ASM Regulation of GPCR signaling Signaling by GPCRs is a highly regulated process One critical way in ... recently 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 ... [39,226–231] Gs Gq RLXN, Cyt, GI CXN CLT1R ET-A/B EDG 1–7 EP2 H1 histamine IP Prostacyclin [232–236] [237–242] [38,243–245] [20,246,247] [248,249] [41,250] Gq Gq Gq, Gi, G1 2/13 Gs Gq Gs CXN, GP CXN, GP GS,...

Ngày tải lên: 13/08/2014, 13:20

... modeling of GPCRs at different activation states, and docking odorant ligands II WEB-BASED SERVER FOR AUTOMATICALLY HOMOLOGY MODELING AND LIGAND DOCKING Homology modeling is process consisting of ... choice for identifying and aligning templates for homology modeling [14] These 44 TP DUY, A GIORGETTI, NHH CHUONG, P CARLONI, H ZUNG methods are used by many of the best protein structure prediction ... 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...

Ngày tải lên: 31/10/2015, 10:39

... following primers: M2-goa1-s, 5¢-CATTATAAGA ACATAGGCGCTACAAGGATGGGTTGTACCATGTC ACAGGAAG-3¢; M2-goa1-PstI-as, 5¢-CCAATGCATTGG TTCTGCAGTTAATACAAGCCGCATCCACGAAGA-3¢ (An engineered PstI recognition ... 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 ... 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 Ga activation...

Ngày tải lên: 07/03/2014, 11:20

... (nucleotides 1–26, sense, initiating ATG underlined) in combination with antisense primers introducing a stop codon sequence: mGRK6-A M2: P2, 5¢-CTAGTCGC TGGAGTTCCCAGAGGAATCTTGGCG-3¢ (nucleotides 1677–1706, ... P2, 5¢-ACTGTCGCTGGAGTTCCCAGAGGAATCTTGG CG-3¢ (nucleotides 1677–1709, antisense) Complementary DNAs encoding the C-terminal mGRK6-A mutants M2 and M3 were prepared either from the mGRK6-A mutant ... (Stratagene, La Jolla, CA) (18 cycles: 94 °C for min, 55 °C for min, 72 °C for min, followed by a single incubation at 72 °C for 10 min) using primer P1, 5¢-CGCCA AGATTCCTCTGGGAACTCCAGCGACAGT-3¢...

Ngày tải lên: 07/03/2014, 12:20

... 35, 231–235 Regulation of Akt signaling pathways by GPCRs New DC & Wong YH (2007) Molecular mechanisms mediating the G protein- coupled regulation of cell cycle progression J Mol Signal 2, Stambolic ... either through matrix metalloproteinases (Gi ⁄ o-, Gq-, and Gs -coupled receptors) or through Rho ⁄ Rho-associated kinase (Rock)-mediated expression of RTK ligands (G1 2 ⁄ 13 -coupled receptors) ... heterotrimeric G protein pathways independently of GPCR activation [41] Akt mediation of GPCR- induced cell cycle control GPCRs have been widely reported to mediate mitogenic signals leading to cellular...

Ngày tải lên: 16/03/2014, 05:20

... following primers were used for cloning at the pLEGFP-N1 vector: GPR30-forward 5¢-TAATAAGTCGACGGGTC TCTTCCT-3¢ and GPR30-reverse 5¢-ATTATTGGATC CTACACGGCACTGC-3¢ Viruses capable of introducing pLEGFP-N1, ... primer 5¢-ATCTCCAAGGCAA GATCA-3¢ and reverse primer 5¢-GTGCCATCAGA CAAGGAA-3¢ were used Primer pair resulted in a PCR product of 216 bp Additionally, forward primer 5¢-GAGCCCCAAGAAGAAAGA-3¢ and reverse ... glucocorticoid The role of GPR in glucocorticoid-mediated signaling was suggested by Schmidt and coworkers [12] b2-adrenergic receptors were able to modulate GR transactivation by stimulating...

Ngày tải lên: 16/03/2014, 18:20

... of 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 ... of devising experiments to observe the effect of domain swapping when both receptors are functional FEBS Journal 272 (2005) 2939–2946 ª 2005 FEBS H GPCR H GPCR GPCR H GPCR H GPCR H GPCR β-arrestin ... ERK1 ⁄ signaling is shown in Fig Regardless of the mechanism by which b-arrestins bind to GPCRs, the signaling pathway activated by these proteins is another way by which GPCR heterooligomerization...

Ngày tải lên: 16/03/2014, 22:20

... Vriend G (2003) Sequence analysis reveals how G protein- coupled receptors transduce the signal to the G protein Proteins 52, 553–560 Madabushi S, Gross AK, Philippi A, Meng EC, Wensel TG & Lichtarge ... Dimerization of G- protein- coupled receptors J Med Chem 44, 4595–4614 Moller S, Vilo J & Croning MD (2001) Prediction of the coupling specificity of G protein coupled receptors to their G proteins Bioinformatics ... structure of G- protein- coupled receptors J Med Chem 40, 3871–3886 Gouldson PR, Snell CR, Bywater RP, Higgs C & Reynolds CA (1998) Domain swapping in G- protein coupled receptor dimers Prot Eng 11, 1181–1193...

Ngày tải lên: 16/03/2014, 22:20

... venom, mimics receptors by activating GTP-binding regulatory proteins (G proteins) J Biol Chem 263, 6491–6494 19 Haga, K., Ogawa, H., Haga, T & Murofushi, H (1998) GTPbinding -protein- coupled receptor ... EQSGYHVEQEKENKLLCEDLPGTEDFVGHQGTVPSDNIDSQGRNCSTNDSLL-OH EPAPAGPRDTDALDLEESSSSDHAERPPGPRRPERGPRGKGKARA EEKESSNDSTSVSAVASNMRDDEITQDENTVSTSLGHSKDENSK KSWKPSAEQMDQDHSSSDSWNNNDAAASLENSASSDEEDIGSETR ... suggested to mediate the internalization of b-adrenergic receptors [45] The GRK-mediated phosphorylation of tubulin may affect physiological processes including GPCRs, and the interaction of GRK2...

Ngày tải lên: 17/03/2014, 09:20

... following primers (Amersham Pharmacia Biotech) were used for PCR: GPR30-forward, 5¢-AGTCGG ATGTGAGGTTCAG-3¢; GPR30-reverse, 5¢-TCTGTGT GAGGAGTGCAAG-3¢; TBP-forward, 5¢-TTTGGAAG AGCAACAAAGG-3¢; ... correlated with growth inhibition The potential importance of the G- proteins in progestin-mediated signaling is also highlighted by a study showing that half of the progesteroneregulated genes were ... AGCAACAAAGG-3¢; TBP-reverse, 5¢-AAGGGTGCAG TTGTGAGAG-3¢ TBP was used to normalize the RNA samples These primer pairs result in PCR products of 240 bp (GPR30) and 243 bp (TBP) LightCycler data were quantitatively...

Ngày tải lên: 24/03/2014, 00:21

... and EcoRI digested pcDNA3 (Invitrogen) using T4 DNA Ligase (Invitrogen) pBABE-GFP and pBABE-GFP-vGPCR were propagated in Escherichia coli strain STBL2 (Invitrogen), whereas pcDNA3-vGPCR, pcDNA3, ... had no significant effect on PGE2 secretion of vGPCRexpressing HUVEC (Fig 5B) These results demonstrate that the increased production of PGE2 in vGPCR-expressing HUVEC is dependent on vGPCR-induced ... KSHV G- protein- coupled receptor (vGPCR) is a constitutively active lytic phase protein with significant homology to the human interleukin-8 (IL-8) receptor and has angiogenic and tumorigenic...

Ngày tải lên: 18/06/2014, 18:20

... and EcoRI digested pcDNA3 (Invitrogen) using T4 DNA Ligase (Invitrogen) pBABE-GFP and pBABE-GFP-vGPCR were propagated in Escherichia coli strain STBL2 (Invitrogen), whereas pcDNA3-vGPCR, pcDNA3, ... had no significant effect on PGE2 secretion of vGPCRexpressing HUVEC (Fig 5B) These results demonstrate that the increased production of PGE2 in vGPCR-expressing HUVEC is dependent on vGPCR-induced ... KSHV G- protein- coupled receptor (vGPCR) is a constitutively active lytic phase protein with significant homology to the human interleukin-8 (IL-8) receptor and has angiogenic and tumorigenic...

Ngày tải lên: 20/06/2014, 01:20

... TPIAESIAVYVFGALIYASKIPERWYPGCFDYFGGSHNLWHLAVLGGIVFHYIAMQE GLSASGFLPIFQIWLTRGGMSVWEHY SPILESLFVYFLGALVYASKVPERWCPGMFDYVGGSHNLWHMAVLGGILFHYNAMQE GLGASLFLPLAHGLSVLGWKRLDAAMGLESFLGLAAINFSGSAVYAMRIPERWFPGTFDLIGQSHNWMHVLVLTGALVRLNGLIR ... EFSFWGQIYGYLSAILYLGSRLPQLLLNFRRKSTEGVSMLFFLFACLGNLTYVLSILAYDGS -SECAAGPGDCEDGEPGQ -EFNILGQVFGWLCAVLYLGSRVPQILLNYRRKSTEGVSMLFFLFACLGNLTYVLSIFAFEPRCRDKHSGIGPHAGGCVGGEAGR SQEPQAVIGMILGYFSAVCYLCARIPQIIKNYREKSCEGLALLFFLLSLTGNLTYGASVIAY ... GLGASLFLPLAHGLSVLGWKRLDAAMGLESFLGLAAINFSGSAVYAMRIPERWFPGTFDLIGQSHNWMHVLVLTGALVRLNGLIR GLGLSGVVPTMHFTIAEGFVKATTVGQMGWFFLMAVMYITGAGLYAARIPERFFPGKFDIWFQSHQIFHVLVVAAAFVHFYGVSN GLGLSGVVPVVHAVGEDGFAALDERMGLKWVMLQGAMYIFGAFIYAARWPERSFPGKFDIWCSSHQIFHIFVLLAAASHLYGMIK...

Ngày tải lên: 14/08/2014, 14:21

... muRGS1 5¢-TCTGCTAGCCCAAAGGATTC-3¢ (sense), 5¢TTCACGTCCATTCCAAAAGTC-3¢ (anti-sense); muRGS2 5¢-GAGAAAATGAAGCGGACACTCT-3¢ (sense), 5¢-TTG CCAGTTTTGGGCTTC-3¢ (antisense); muHPRT as housekeeping gene ... control A Stimulation h Gene Control relative fluorescence LPS relative fluorescence (fold) RGS1 RGS2 RGS3 RGS4a RGS5 RGS6a RGS7a RGS9a RGS10 RGS11a RGS14 RGS16 RGS17a RGS18 RGS19 RGS20a 168 1532 153 ... dominant-negative Gai protein constructs [11] G proteins are located downstream of G- protein- coupled receptors (GPCR) [12] GPCR represent a large family of cell-surface proteins mediating the effects...

Ngày tải lên: 18/02/2014, 13:20

... by G protein- gated inwardly rectifying potassium (GIRK) channels Pflugers Arch 456, 1097– 1108 Stringer BK, Cooper AG & Shepard SB (2001) Overexpression of the G- protein inwardly rectifying potassium ... signalling molecules) GIRK1, GIRK4, Gb ⁄ c, PKA, PP2A and protein phosphatase was identified to exist in rat atrial membranes in situ [21], supporting the physiological relevance of this regulation ... heterooligomeric combinations, including not only GIRK1 ⁄ GIRK4, but also the homooligomeric GIRK1 subunit alone, have been identified to be under PKA regulation [18] In addition, the GIRK1 protein...

Ngày tải lên: 07/03/2014, 00:20

... effector of G- proteins, its function being governed by whether the G- protein is in an ‘on state’ [being activated by receptors, e .g OA1 G- protein- coupled receptor A (GPCR) ], or in an ‘off state’ In the ... among all the organelles, the Golgi bodies seem to show an abundance of G- proteins, involved in their biogenesis, trafficking, membrane organization, and many other important functions [16–19] G- proteins ... ‘on’ state (GTP-bound state, activated by the ligand-bound GPCR) Ligand activation of the GPCR leads to receptor-mediated activation of the G- protein, in which GTP replaces the bound GDP, and therefore...

Ngày tải lên: 07/03/2014, 00:20

... TTTTGTCGACATGGCGCAACACGATGAAGC CGTAGACAAC GGGGGGATCCTTACATAAGCGTACAACAAA CACTATTTGATTTCGGCGCCTGAGCATCA TTTAGCTTTTT TTTTCTCGAGAAAGATGCCGATTTGGGCGC GGGGCTCGAGGTTTTATATTTGTTGTAAAA GCCCCTCGAGATATTATATATATATATAGG ... GCCCCTCGAGATATTATATATATATATAGG TAAAGGATCCCTTGTCATCGTCATCCTTGT AGTCAACACTATTTGAGTTTGACATTTGGC GAGAGAATTCGGGGGACCGTCAGTCTTCCT CTTCCCCC TTCCGAATTCTCATTTACCCGGAGACAGGG CCCCGCGGCCGCTGACACCGATTATTTAAA TTTTGAGCTCGGAGCCATAATGACAGCAGT ... TTTTGAGCTCGGAGCCATAATGACAGCAGT TTTTGTCGACATGGCGCAACACGATGAAGC CGTAGACAAC GGGGGGATCCTTACATAAGCGTACAACAAA CACTATTTGATTTCGGCGCCTGAGCATCA TTTAGCTTTTT ATCCAAAGTTTAGCCGATGACCCAAGCCAA TTGGCTTGGGTCATCGGCTAAACTTTGGAT AAACGCCTTCGCCCAAAGTTTAAAAGATGA...

Ngày tải lên: 16/03/2014, 01:20