Abstract:
:Although G protein-coupled receptor (GPCR) kinases (GRKs) have been shown to mediate desensitization of numerous GPCRs in studies using cellular expression systems, their function under physiological conditions is less well understood. In the current study, we employed various strategies to assess the effect of inhibiting endogenous GRK2/3 on signaling and function of endogenously expressed G s-coupled receptors in human airway smooth muscle (ASM) cells. GRK2/3 inhibition by expression of a Gbetagamma sequestrant, a GRK2/3 dominant-negative mutant, or siRNA-mediated knockdown increased intracellular cAMP accumulation mediated via beta-agonist stimulation of the beta-2-adrenergic receptor (beta 2AR). Conversely, neither 5'-( N-ethylcarboxamido)-adenosine (NECA; activating the A2b adenosine receptor) nor prostaglandin E2 (PGE 2; activating EP2 or EP4 receptors)-stimulated cAMP was significantly increased by GRK2/3 inhibition. Selective knockdown using siRNA suggested the majority of PGE 2-stimulated cAMP in ASM was mediated by the EP2 receptor. Although a minor role for EP3 receptors in influencing PGE 2-mediated cAMP was determined, the GRK2/3-resistant nature of EP2 receptor signaling in ASM was confirmed using the EP2-selective agonist butaprost. Somewhat surprisingly, GRK2/3 inhibition did not augment the inhibitory effect of the beta-agonist on mitogen-stimulated increases in ASM growth. These findings demonstrate that with respect to G s-coupled receptors in ASM, GRK2/3 selectively attenuates beta 2AR signaling, yet relief of GRK2/3-dependent beta 2AR desensitization does not influence at least one important physiological function of the receptor.
journal_name
Biochemistryjournal_title
Biochemistryauthors
Kong KC,Gandhi U,Martin TJ,Anz CB,Yan H,Misior AM,Pascual RM,Deshpande DA,Penn RBdoi
10.1021/bi801056wsubject
Has Abstractpub_date
2008-09-02 00:00:00pages
9279-88issue
35eissn
0006-2960issn
1520-4995journal_volume
47pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract::Protein kinases are key regulators of diverse signaling networks critical for growth and development. Protein kinase A (PKA) is an important kinase prototype that phosphorylates protein targets at Ser and Thr residues by converting ATP to ADP. Mg(2+) ions play a crucial role in regulating phosphoryl transfer and can l...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi901475g
更新日期:2009-12-08 00:00:00
abstract::Cold-shock proteins (Csps) are expressed at lower-than-optimum temperatures, and they function as RNA chaperones; however, no structural studies on psychrophilic Csps have been reported. Here, we aimed to investigate the structure and dynamics of the Csp of psychrophile Colwellia psychrerythraea 34H, ( Cp-Csp). Althou...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.8b00144
更新日期:2018-07-03 00:00:00
abstract::The reaction of nitric oxide (NO) with fast cytochrome bo from Escherichia coli has been studied by electronic absorption, MCD, and EPR spectroscopy. Titration of the enzyme with NO showed the formation of two distinct species, consistent with NO binding stoichiometries of 1:1 and 2:1 with observed dissociation consta...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi971481a
更新日期:1997-12-23 00:00:00
abstract::The kinetics of self-association for beef liver glutamate dehydrogenase (EC 1.4.1.3) have been measured by using pressure perturbation in both the time domain and the frequency domain by monitoring scattered light intensity. The kinetic behavior is entirely consistent with the random self-association model proposed by...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00579a007
更新日期:1979-06-12 00:00:00
abstract::C-type natriuretic peptide (CNP) binds the guanylyl cyclase-linked natriuretic peptide receptor B (NPR-B) and stimulates marked elevations of the intracellular signaling molecule, cGMP. Here, the essential role of phosphorylation in the hormonal activation and deactivation of this receptor is described. Exposure of NI...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi972303k
更新日期:1998-02-24 00:00:00
abstract::Cell-signaling peptides have been extensively used to transport functional molecules across the plasma membrane into living cells. These peptides consist of a hydrophobic sequence and a cationic nuclear localization sequence (NLS). It has been assumed that the hydrophobic region penetrates the hydrophobic lipid bilaye...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi061895g
更新日期:2007-01-30 00:00:00
abstract::Tropomyosin (TM) is a coiled-coil that binds head-to-tail along the helical actin filament. The ends of 284-residue tropomyosins are believed to overlap by about nine amino acids. The present study investigates the function of the N- and C-terminal overlap regions. Recombinant tropomyosins were produced in Escherichia...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi000242b
更新日期:2000-06-13 00:00:00
abstract::Formation of protein-protein assemblies is essential in maintaining cell structure and function. Conservation of structural motifs and binding sites is the result of evolutionary pressure for solutions compatible with both molecular economy and regulation. PDZ domains are a typical example: A conserved fold governs sp...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi061869i
更新日期:2007-01-30 00:00:00
abstract::In the presence of Ca2+ and calmodulin (CM), purified smooth muscle myosin light chain kinase (MLCKase) was found to undergo autophosphorylation at a rate that was about 200-fold slower than its catalytic activity. Up to 1.7 mol of phosphate were incorporated per mole of kinase. Lower levels of incorporation could be ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00037a025
更新日期:1995-09-19 00:00:00
abstract::The peptidolytic reaction of HIV-1 protease has been investigated by using four oligopeptide substrates, Ac-Ser-Gln-Asn-Tyr-Pro-Val-Val-NH2, Ac-Arg-Ala-Ser-Gln-Asn-Tyr-Pro-Val-Val-NH2, Ac-Ser-Gln-Ser-Tyr-Pro-Val-Val-NH2, and Ac-Arg-Lys-Ile-Leu-Phe-Leu-Asp-Gly-NH2, that resemble two cleavage sites found within the natu...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00098a023
更新日期:1991-08-27 00:00:00
abstract::Drug intervention that prevents reabsorption of circulating bile acids by the apical (ileal) sodium/bile acid cotransporter (ASBT) may be a promising new therapy for lowering of plasma cholesterol. 2164U90 is a benzothiazepine-based competitive inhibitor of bile acid transport with K(i) values of approximately 10 and ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0205404
更新日期:2002-12-17 00:00:00
abstract::The bacterial reaction center (RC) converts light into chemical energy through the reduction of an internal quinone molecule Q(B) to Q(B)H(2). In the native RC, proton transfer is coupled to electron transfer and is not rate-controlling. Consequently, proton transfer is not directly observable, and its rate was unknow...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi020419x
更新日期:2002-12-17 00:00:00
abstract::Phage display technique is a powerful tool with which to identify novel binding sequences for antibody and receptor targets. Few studies, however, have used this technology to select affinity peptides for ligand molecules. Here, we screened a peptide phage library for binding to toxic shock syndrome toxin 1 (TSST-1) t...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi960132y
更新日期:1996-08-13 00:00:00
abstract::We studied hyperosmolarity-induced changes in cell volume and cytoplasmic pH in PS120 cells expressing Na(+)/H(+) exchanger (NHE) isoforms and their mutants. Change in cell volume was estimated by measuring change in cell height by means of confocal microscopy. Regulatory volume increase (RVI) and cytoplasmic alkalini...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi020427d
更新日期:2003-02-04 00:00:00
abstract::Four different temperature-sensitive M protein mutants (tsM) of vesicular stomatitis virus (VSV) were characterized with regard to the association of the mutated M protein either with nucleocapsids or with membranes in the intact virions. Virions were labeled with the photoreactive hydrophobic probe [125I]iodonaphthyl...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00527a020
更新日期:1981-11-24 00:00:00
abstract::The bond cleavage step of pepsinogen activation has been investigated in a kinetic study in which the denatured products of short-term acidifications were separated on SDS-polyacrylamide gels and the peptide products were quantitated by densitometry. Although several peptide products were observed, under the condition...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00432a040
更新日期:1989-03-21 00:00:00
abstract::A detailed kinetic analysis of the catalytic trimer of aspartate transcarbamoylase containing the active site substitution H134A was performed to investigate the role of His 134 in the catalytic mechanism. Replacement of histidine by alanine resulted in decreases in the affinities for the two substrates, carbamoyl pho...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00143a031
更新日期:1992-07-21 00:00:00
abstract::How various anti-amyloidogenic compounds inhibit the formation of Alzheimer's beta-amyloid fibrils (fAbeta) from amyloid beta-peptide (Abeta) and destabilize fAbeta remains poorly understood. Using spectrophotometry, spectrofluorometry, atomic force microscopy, sodium dodecyl sulfate-polyacrylamide gel electrophoresis...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi061540x
更新日期:2007-02-20 00:00:00
abstract::The interaction of linker histone H1 with both linear and superhelical double-stranded DNA has been investigated at low ionic strengths. Gel mobility retardation experiments demonstrate strikingly different behavior for the two forms of DNA. First, the experiments strongly suggest that linker histone binds to superhel...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0497704
更新日期:2004-06-22 00:00:00
abstract::The reaction of enantiomerically pure (2S)-N-acetyl-L-alanyl-L-phenylalanyl alpha-chloroethane with gamma-chymotrypsin was studied as a probe of the mechanism of inactivation of serine proteases by peptidyl chloroalkanes. It was determined crystallographically that the peptidyl chloroethane alkylates His57 with retent...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00250a033
更新日期:1994-11-22 00:00:00
abstract::The Rhodnius prolixus aggregation inhibitor 1 (RPAI-1) is a novel blood-sucking salivary molecule that binds to ADP and attenuates platelet aggregation. In this report, we determine the binding constants and specificity of RPAI-1 for adenine nucleotides and its functional significance. By the Hummel-Dreyer method of e...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi011015s
更新日期:2002-03-19 00:00:00
abstract::Rh(phen)2phi3+ (phen = 1,10-phenanthroline; phi = 9,10-phenanthrenequinone diimine) avidly binds to DNA via intercalation from the major groove and upon photoactivation produces strand scission with single base 5' asymmetry. Enantiomers of Rh(phen)2phi3+, which lack hydrogen-bonding substituents in ancillary positions...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00180a005
更新日期:1994-04-12 00:00:00
abstract::Mutations in the Kirsten ras (KRAS) gene are present in almost all pancreatic adenocarcinomas, and one common mutation is at codon 12: GGT (Gly) is transformed into GAT (Asp). In this work we have targeted the KRAS coding sequence embracing the GAT mutation with a sense PNA molecule (P14), with the aim of downregulati...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0505215
更新日期:2005-08-09 00:00:00
abstract::Interaction between aggregates of amyloid beta protein (Abeta) and membranes has been hypothesized by many to be a key event in the mechanism of neurotoxicity associated with Alzheimer's disease (AD). Proposed membrane-related mechanisms of neurotoxicity include ion channel formation, membrane disruption, changes in m...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9006397
更新日期:2009-09-22 00:00:00
abstract::The aromatic hydroxylation of six pairs of selectively deuterated monosubstituted benzenes was investigated with rat liver microsomes of various induction states. The substrates studied included 3,5-D2C6H3X (1a-6a) and 2,4,6-D3C6H2X (1b-6b), where X = Br, CN, NO2, OCH3, CH3, or Ph, respectively. The deuterium content ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00308a031
更新日期:1984-06-19 00:00:00
abstract::The gene for human pancreatic cholesterol esterase consists of 11 exons and 10 introns and is 9.2 kb in length. The last and longest exon (841 nucleotides) is unique to the human gene. Functional amino acids are encoded on separate exons. The leader sequence is encoded by a single exon which carries two additional N-t...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00141a017
更新日期:1992-07-07 00:00:00
abstract::Transcripts of Saccharomyces cerevisiae nuclear tRNA genes are normally terminated within a few nucleotides of the tRNA coding region, in contrast to mitochondrially encoded tRNAs, which are contained within polycistronic transcripts and thus require 3'-processing by mitochondrial endonucleases. We show that 3'-proces...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00159a024
更新日期:1992-11-10 00:00:00
abstract::S-(4-Bromo-2,3-dioxobutyl)glutathione (S-BDB-G), a reactive analogue of glutathione, has been synthesized and characterized by UV spectroscopy and thin-layer chromatography, as well as by bromide and primary amine analysis. Incubation of S-BDB-G (200 microM) with the 4-4 isoenzyme of rat liver glutathione S-transferas...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00111a006
更新日期:1991-11-26 00:00:00
abstract::The HIV-1 fusion peptide serves as a useful model system for understanding viral/target cell fusion, at least to the lipid-mixing stage. Previous solid-state NMR studies have shown that the membrane-bound HIV-1 fusion peptide adopts an extended conformation in a lipid mixture close to that of host cells of the virus. ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0348157
更新日期:2003-10-14 00:00:00
abstract::The molecular chaperones Hsp90 and Hsp70 are highly regulated by various cochaperones that participate in the activation of steroid receptors. Here we study Tpr2 (also called DjC7), a TPR domain-containing type III J protein implicated in steroid receptor chaperoning. We propose that Tpr2 plays a role in the Hsp90-dep...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi800770g
更新日期:2008-08-05 00:00:00