functional expression of g protein coupled receptors in yeast

... 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, ... homologous GPCR desensitization in the airway This may preferentially affect β2AR signaling in ASM, in light of findings by McGraw et al suggesting that low (endogenous) expression levels of GRKs in ... increased by sloughing of airway epithelium Second, Gq- or Gi -coupled receptor signaling may be sensitized, or Gs -coupled receptor signaling desensitized, resulting in an imbalance of signaling...

Ngày tải lên: 13/08/2014, 13:20

... 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 ... The increase of GTPcS binding was completely inhibited by Fig Binding of muscarinic ligands All experiments were performed in triplicate Each data point represents the mean ± SEM (A) Binding of...

Ngày tải lên: 07/03/2014, 11:20

... 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 ... predominantly in ‘single point’ assays In these, single amounts of Renilla luciferase and fluorescent protein- tagged forms of a single GPCR, to study homodimerization ⁄ oligomerization, or pairs of ... that although single point mutations in both transmembrane region III and transmembrane region VI prevented binding of antagonist radioligands (including [3H]mepyramine), coexpression of the two...

Ngày tải lên: 16/03/2014, 22:20

... 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 ... surface: a short chain containing an ethylenediamine spacer in and 5, and a long chain containing a PEG spacer in and In conjugates and 6, there was an optional cofunctionalization of the QD surface...

Ngày tải lên: 11/08/2014, 00:22

... mouse 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 ... SERVER FOR AUTOMATICALLY HOMOLOGY MODELING AND LIGAND DOCKING Homology modeling is process consisting of main steps: template search, targettemplate sequence alignment, model construction and ... classes of model generation methods have been proposed: (i) Modeling by assembly of rigid bodies; (ii) Modeling by segment matching or coordinate reconstruction; (iii) Modeling by satisfaction of...

Ngày tải lên: 31/10/2015, 10:39

... agonist binding domain and the G- protein coupling domain are part of the same subunit Coupling between ligand binding and HD activation has also been recently examined in the homodimeric mGlu receptors ... 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 composed of these ... receptors, the 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...

Ngày tải lên: 07/03/2014, 21:20

... 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 ... (1999) G- protein coupled ¨ receptor kinases as modulators of G- protein signalling J Physiol 517, 5–23 Ferguson SS (2001) Evolving concepts in G proteincoupled receptor endocytosis: the role in receptor ... 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...

Ngày tải lên: 07/03/2014, 12:20

... chemokine CCR2b and CCR5 receptors that gained coupling selectivity for G1 1 -protein in cotransfected HEK-293 cells [14]; (c) dopamine D1 and D2 receptors that gained coupling selectivity for Gq-proteins ... role in GPCR activation of mitogen-activated protein kinase (MAPK) In this context it may act as an adaptor or scaffolding for recruiting signaling molecules into a complex along with the agonist-occupied ... 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...

Ngày tải lên: 16/03/2014, 22:20

... 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 ... 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 ... reasonable insights into the structural and functional features of the aminergic receptors Among them, 16 of 33 locations that are known to be involved in ligand binding in aminergic receptors...

Ngày tải lên: 16/03/2014, 22:20

... primers The fluorogenic probes were CCGCTGAGCCTCGTCCATGGG, TLR2; TTCGTTCAACTTCCACCAAGAGCTGCCT, TLR4; TACACACAATATATGTCTGCAGGAGGCCCA, TLR5; and AGCACCCTCAACTTCACCTTGGATCTGTC, TLR A GeneAmp 5700 Sequence ... TLR5; and TCTGAAGACTTCAGGCCCAACT, TLR9 for forward primers, and GCAGCTCTCAGATTTACCCAAAA, TLR2; TTATCTGAAGGTGTTGCACATTCC, TLR4; TTAAGACTTCCTCTTCATCACAACCTT, TLR5; and TGCACGGTCACCAGGTTGT, TLR9 for ... Technologies), and 3.575 μl DNAase free water The oligonucletoide primers for respective TLRs were CTGCAAGCTGCGGAAGATAAT, TLR2; AGAGTTTCCTGCAATGGATCAAG, TLR4; GGCTTAATCACACCAATGTCACTATAG, TLR5;...

Ngày tải lên: 08/08/2014, 16:23

... ttcggcatcagctttgtgctctgtatctcgtgcatcttggtgaagaccaatcgcgtcctcctggtatttgaagccaagatacccaccagcttc caccggaagtggtgggggctcaacct gcagttcctgctggttttcctctgcaccttcatgcagatcctcatctgcatcatctggctctacacggcgcccccctc tagcaccgcaaccatgagctggaagacgaaatcatcttca The sequence ... detected, indicating the tested RNA samples were free of genomic DNA contamination Sequencing results were as follows: ttcggcatcagctttgtgctctgtatctcgtgcatcttggtgaagaccaatcgcgtcctcctggtatttgaagccaagatacccaccagcttc ... sense 5’-ttcggcatcagctttgtg-3’, antisense 5’-tgaagatgatttcgtcttcc3’; (2) GAPDH: sense 5’-ctcaactacatggtctacatg -3’, antisense 5’-tggcatggactgtggtcatgag-3’, yielding predicted products of 234 and...

Ngày tải lên: 10/08/2014, 05:21

... immunofluorescence histochemical staining to detect estrogen-binding activity and found of tumors had estrogen-binding activity and the intensity of the staining was found to be much greater in the tumors from ... and that of 88 Figure sion of progesterone receptors Photomicrograph showing breast tissue with strong expresPhotomicrograph showing breast tissue with strong expression of progesterone receptors ... (Figure 1) Expression of estrogen and progesterone receptors and their potential role in the progression of vestibular schwannoma is still an area of controversy Many diverging studies, using various...

Ngày tải lên: 11/08/2014, 07:21

... TCATCTCTGAAGAGGATCTG-3¢) and (5¢-GCATG CCTGCAGGTCGACTCTAGAGGATCTCAAGCCAGT GACCGCCTCCC-3¢); for Golf (5¢-GGTACCGCTGCAA TGGGGTGTTTGGGCAAC-3¢) and (5¢-GCGGCCGCCT CAGATCACAAGAGTTCGTACTGC-3¢); for Ga15 ... (5¢-CGTCAAGGAGAAAAAACCCCGGATCT AAAAAATGGAGCGAAGGAACCACAG-3¢) and (5¢-AG CTGCCTGCAGGTCGACTCTAGAGGATCCTAACCAA TTTTGCTGCC-3¢); for OR17-40 (5¢-CGTCAAGGAG AAAAAACCCCGGATCTAAAAAATGGAGCAGAAAC TCATCTCTGAAGAGGATCTG-3¢) ... CCCGGATCTAAAAAATGGAGCAGAAACTCATCTC TGAAGAGGATCTG-3¢) and (5¢-GCATGCCTGCAGG TCGACTCTAGAGGATCTCAAGCCAGTGACCGCCT CCC-3¢), and checked for the presence and sequence of the new insert, as in the case of pJH2-I7 Plasmids...

Ngày tải lên: 07/03/2014, 16:20

... 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, ... using the t-test as indicated in Fig C, control GPR30 C TIF-2 C GPR30 Fig GPR30 down-regulates the expression of TIF2 protein The regulation of cofactor was studied in HME cells stably expressing ... suggesting the antiproliferative activity of GPRs [21–23] The role of GPR30 in glucocorticoid-mediated signaling has not hitherto been described Many groups independently demonstrated GPR30 in...

Ngày tải lên: 16/03/2014, 18:20

... Kameyama, K., Haga, K., Haga, T., Moro, O & Sadee, W (1994) Activation of a GTP-binding protein and a GTP-binding-proteincoupled receptor kinase (b-adrenergic-receptor kinase-1) by a muscarinic receptor ... fusion proteins of full-length bI-tubulin, bIII-tubulin, and C-terminal peptides of bI-tubulin expressed in E coli We cloned the bI-tubulin and bIII-tubulin genes GST fusion proteins of bI-tubulin ... fusion protein with GST of a peptide corresponding to positions 393–445 of rat bI-tubulin was also expressed in E coli using an expression vector, pGEX4T-3 (GST-b-tubulinC) These GST fusion proteins...

Ngày tải lên: 17/03/2014, 09:20

... [4] In KS, KSHV is found in spindle cells at all stages of the disease [1] Changes in endothelial gene expression resulting from KSHV encoded gene products could provide insights into pathogenesis ... selective COX-1 inhibitor SC560 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 ... resulting biosynthesis of PGE2 are sufficient to stimulate all of the key features of carcinogenesis including mutagenesis, mitogenesis, angiogenesis, metastasis, inhibition of apoptosis and immunosuppression...

Ngày tải lên: 18/06/2014, 18:20

... [4] In KS, KSHV is found in spindle cells at all stages of the disease [1] Changes in endothelial gene expression resulting from KSHV encoded gene products could provide insights into pathogenesis ... selective COX-1 inhibitor SC560 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 ... resulting biosynthesis of PGE2 are sufficient to stimulate all of the key features of carcinogenesis including mutagenesis, mitogenesis, angiogenesis, metastasis, inhibition of apoptosis and immunosuppression...

Ngày tải lên: 20/06/2014, 01:20

... was any staining of the nucleus, regardless of staining intensity Cell counting was performed in six high-power microscopic fields (400x, Standard 20, ZEISS), which comprised almost all of the OLP ... reported that 5% of the cases of oral squamous cell carcinoma show diminished expression of hMSH2 protein In the present study, we demonstrated decreased expression of hMSH2 protein in reticular ... important in distinguishing and repairing misparing and slippage errors in DNA synthesis The mismatch is detected by two complexes, the hMSH2-GTBP/hMSH6 and the hMSH2-hMSH3 heterodimers after interaction...

Ngày tải lên: 03/11/2012, 09:54

... Quercetin-4¢-Glc (spiraeoside) Quercetin-7-Glc Quercetin-3,4¢-diGlc Flavones Apigenin-7-Glc (apigetrin) Luteolin-4¢-Glc Luteolin-7-Glc Luteolin-3¢,7-diGlc Flavanones Naringenin-7-Glc Eriodictyol-7-Glc ... Kaempferol-3-Glc Isorhamnetin-3-Glc Naringenin-7-GlcRha (naringin) Hesperetin-7-GlcRha (hesperidin) Phloretin-7-Glc (phlorizin) Oleuropein Cyanidin-3-Glc (kuromanin) Cyanidin-3,5-diGlc Flavanones ... incubating for 30 at 37 °C, and measuring the release of 4NP Protein assays and protein sequencing Total protein in crude and semipuri®ed samples was estimated using the Pierce Protein Assay Reagent...

Ngày tải lên: 08/03/2014, 16:20

... Murga C, Laguinge L, Wetzker R, Cuadrado A & Gutkind JS (1998) Activation of Akt ⁄ protein kinase B by G protein- coupled receptors A role for a and bc subunits of heterotrimeric G proteins acting ... aspects of cellular signaling, much remains to be uncovered in order to deepen our understanding of how individual GPCRs functionally interact with different G proteins to initiate a cascade of events ... G- protein bc-dimers in growth and differentiation Oncogene 20, 1653–1660 20 Murga C, Fukuhara S & Gutkind JS (2000) A novel role for phosphatidylinositol 3-kinase b in signaling from G protein- coupled...

Ngày tải lên: 16/03/2014, 05:20