g protein coupled receptors and downstream effectors

... concentration than GDP This exchange of the G- protein- bound GDP for GTP induces a conformational change in the switch region of G and causes the dissociation of G from the G γ dimer The G and G subunits ... [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, ... 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...

Ngày tải lên: 13/08/2014, 13:20

... modeling of GPCRs at different activation states automatic modeling of GPCRs at different activation states, and docking odorant ligands II WEB-BASED SERVER FOR AUTOMATICALLY HOMOLOGY MODELING AND ... the ligands AutoDock Vina is a new open-source program for drug discovery, molecular docking and virtual screening, offering multi-core capability, high performance and enhanced accuracy and ease ... CHARACTERIZATION OF G PROTEIN COUPLED RECEPTORS 47 Fig The lowest Autodock VINA affinity of docked ligands generated by the receptor built by template 3rze, although their DOPE and GA341 scores are...

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

... 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 ... 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 ... 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 and be trafficked...

Ngày tải lên: 07/03/2014, 21:20

... (2002) G- proteincoupled receptor oligomerization and its potential for drug discovery Nature Rev Drug Discovery 1, 808–820 12 Milligan G (2004) G protein- coupled receptor dimerization: function and ... signal was obtained with increasing energy acceptor ⁄ energy donor ratios Saturation curves so generated resemble ligand ⁄ GPCR binding curves and, as such, the ratio of energy acceptor ⁄ energy ... (2000) Oligomerization of l- and d-opioid receptors J Biol Chem 275, 26128–26135 57 Devi LA (2001) Heterodimerization of G- protein- coupled receptors: pharmacology, signaling and trafficking Trends...

Ngày tải lên: 16/03/2014, 22:20

... native ligand is a small molecule Previously, peptide ligands and small neurotransmitter-like molecules were coupled to QDs resulting in specific interactions with the target receptors and drug transporters ... using covalently tethered nucleoside agonist ligands Various approaches to the linking chemistry and the nature of the spacer group and solubilizing groups were compared The QD nucleoside conjugates ... affinity depended greatly on the type of coating and covalent linkage to the QD Conjugates tethered as monovalent attachments through amide-linked chains and PEG dis- Figure General design features...

Ngày tải lên: 11/08/2014, 00:22

... 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) ... activation G1 2 ⁄ 13-, Gi ⁄ o-, Gq-, and Gs -coupled receptors are all known to activate the phosphoinositide 3-kinase (PI3K) ⁄ Akt pathway through either Ga or Gbc subunits Gaq and Gas subunits ... depression and pain [18] GPCRs preferentially couple to heterotrimeric G proteins (consisting of a, b and c subunits) that are grouped into four classes, known as Gaq ⁄ 11, Gai ⁄ o, Gas and Ga12 ⁄...

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, ... 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 ... or anti-b-actin (Sigma) Peroxidase-conjugated goat anti-rabbit IgG (Cappel, West Chester, PA, USA) for GR and peroxidase-conjugated goat anti-mouse IgG (Cappel) for PR, ER and b-actin were used...

Ngày tải lên: 16/03/2014, 18:20

... may form hetero-oligomers [5] The same phenomenon has been shown to occur in hetero-oligomers formed by Gi -coupled and Gq -coupled receptors [6] and by Gs -coupled and Gq -coupled receptors [7] One ... activate a G- protein, leaving their coupling efficacy unchanged For instance, b2-adrenergic receptors, which are coupled with stimulatory G- proteins, and d-opioid and j-opioid receptors, which are coupled ... 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...

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 ... 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 ... 1181–1193 Gouldson PR, Higgs C, Smith RE, Dean MK, Gkoutos GV & Reynolds CA (2000) Dimerization and domain swapping in G- protein- coupled receptors: a computational study Neuropsychopharmacology 23,...

Ngày tải lên: 16/03/2014, 22:20

... 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 ... (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, ... proteins were visualized by SDS ⁄ PAGE and autoradiography of the gel Miscellaneous Protein concentrations were determined according to Bradford [44] using bovine IgG as standard SDS ⁄ PAGE and...

Ngày tải lên: 07/03/2014, 12:20

... 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 ... KSWKPSAEQMDQDHSSSDSWNNNDAAASLENSASSDEEDIGSETR KAFLHWYTGEGMDEMEFTEAESNMNDLVSEYQQYQDATADEEGEMY EDDDEESEAQGPK-OH NEEGAPQEGILEDMPVDPDNEAYEMPSEEGYQDYEPEA-OH PEEVAQEAAEEPLIEPLMEPEGESYEDPPQEEYQEYEPEA-OH EQFAEEFFAADVESFLNEYGLLPEREIHDLGQTNTEDEDIE-OH...

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¢; ... 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 ... tissues [12–14], and some G protein- coupled receptors are known to be involved in growth regulation The regulation of GPR30 expression in MCF-7 cells was progestin specific and was not upregulated by...

Ngày tải lên: 24/03/2014, 00:21

... regulate the activity of downstream effectors GTP on G is hydrolyzed to GDP by its own GTPase activity as well as GTPase-activating proteins, such as regulator of G protein signaling (RGS) proteins ... phosphate (cGMP) phosphodiesterase, and so forth); Gq family (Gqα, G1 1α, G1 4α, G1 5α, and G1 6α; stimulate phospholipase C-β isozymes); and G1 2 subfamily (G1 2α and G1 3α; regulate Rho guanine nucleotide ... 17 genes that encode G protein α subunits have been identified and they can be grouped into four subfamilies: Gs, Gi, Gq, and G1 2/13 Significant advances in our understanding of the structure and...

Ngày tải lên: 10/04/2014, 22:24

... 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, and ... Virology, Baltimore, MD) by BglII and EcoRI digestion [12] The digested vGPCR fragment was separated by gel electrophoresis, purified using QIAquick PCR Purification Kit (Qiagen, Page of (page number ... 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

... 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, and ... Virology, Baltimore, MD) by BglII and EcoRI digestion [12] The digested vGPCR fragment was separated by gel electrophoresis, purified using QIAquick PCR Purification Kit (Qiagen, Page of (page number ... 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

... MasterPlus SYBR Green I mix (Roche Diagnostics) The following primers were used: muRGS1 5¢-TCTGCTAGCCCAAAGGATTC-3¢ (sense), 5¢TTCACGTCCATTCCAAAAGTC-3¢ (anti-sense); muRGS2 5¢-GAGAAAATGAAGCGGACACTCT-3¢ ... 5¢-GAGAAAATGAAGCGGACACTCT-3¢ (sense), 5¢-TTG CCAGTTTTGGGCTTC-3¢ (antisense); muHPRT as housekeeping gene 5¢-ACTTTGCTTTCCCTGGTTA-3¢ (sense), 5¢-CAAAGTCTGGCCTGTATCC-3¢ (antisense); muTNF-a 5¢-GACCCTCACACTCAGATCATCTTC-3¢ (sense), ... 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

... [16–19] G- proteins on the Golgi membranes also engage in a plethora of very specific protein protein interactions, recognizing downstream effectors [20–22] Understanding the origins of these specificities ... 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 ... calcium ions is an emerging and interesting area for investigation In view of these observations, interactions between the Ca2+-binding protein Calnuc and signaling molecule G- proteins assume extreme...

Ngày tải lên: 07/03/2014, 00:20

... Chillakuri et al 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 ... 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 The reverse ... primer for the CB1 gene with the BamHI restriction site was 5¢-GC G GAT CC G ACC ATG GCG AAG TCG ATC CTA GAT GGC-3¢ The reverse primer for the full-length CB1 gene, with EcoRI and NotI restriction...

Ngày tải lên: 23/03/2014, 07:20