Abstract：
:Adenosine A(2B) receptors, which play a role in inflammation and cancer, are of considerable interest as novel drug targets. To gain deeper insights into ligand binding and receptor activation, we exchanged amino acids predicted to be close to the binding pocket. The alanine mutants were stably expressed in CHO cells and characterized by radioligand binding and cAMP assays using three structural classes of ligands: xanthine (antagonist), adenosine, and aminopyridine derivatives (agonists). Asn282(7.45) and His280(7.43) were found to stabilize the binding site by intramolecular hydrogen bond formation as in the related A(2A) receptor subtype. Trp247(6.48), Val250(6.51), and particularly Ser279(7.42) were shown to be important for binding of nucleosidic agonists. Leu81(3.28), Asn186(5.42), and Val250(6.51) were discovered to be crucial for binding of the xanthine-derived antagonist PSB-603. Leu81(3.28), which is not conserved among adenosine receptor subtypes, may be important for the high selectivity of PSB-603. The N186(5.42)A mutant resulted in an increased potency for agonists. The interactions of the non-nucleosidic agonist BAY60-6583 were different from those of the nucleosides: while BAY60-6583 appeared not to interact with Ser279(7.42), its interactions with Trp247(6.48) and Val250(6.51) were significantly weaker compared to those of NECA. Moreover, our results discount the hypothesis of Trp247(6.48) serving as a "toogle switch" because BAY60-6583 was able to activate the corresponding mutant. This study reveals distinct interactions of structurally diverse ligands with the human A(2B) receptor and differences between closely related receptor subtypes (A(2B) and A(2A)). It will contribute to the understanding of G protein-coupled receptor function and advance A(2B) receptor ligand design.
journal_name
Biochemistryjournal_title
Biochemistryauthors
Thimm D,Schiedel AC,Sherbiny FF,Hinz S,Hochheiser K,Bertarelli DC,Maass A,Müller CEdoi
10.1021/bi3012065subject
Has Abstractpub_date
2013-01-29 00:00:00pages
726-40issue
4eissn
0006-2960issn
1520-4995journal_volume
52pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract：:13C NMR spectroscopy has been used to study the amino group environments and metal binding properties of 13C reductively methylated bovine alpha-lactalbumin. Bovine alpha-lactalbumin is a Ca2+ metalloprotein containing 12 lysyl amino groups and a free amino terminus. All 13 amino groups can be 13C-dimethylated without...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00315a026
更新日期：1984-09-25 00:00:00
abstract：:We have sought to determine the role of histone H2A phosphorylation in chromatin by examining the distribution of the phosphorylated and unphosphorylated forms of this core histone within the nuclei of mouse and human cells. At any time, only about 15% of the H2A of whole chromatin is in the phosphorylated form, and i...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00539a021
更新日期：1982-05-11 00:00:00
abstract：:Inosine has the ability to act as a "wild-card" binding nonspecifically to both A.T and G.C base pairs. This has obvious implications for the design of oligonucleotide site-directed probes. In this paper we present a series of oligonucleotides with a 5'pur9-pyr9-pyr9 motif which are designed to fold up sequentially in...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi960193w
更新日期：1996-10-15 00:00:00
abstract：:The cytochrome aa3-type terminal quinol oxidase of Bacillus subtilis catalyzes the four-electron reduction of dioxygen to water. It resembles the aa3-type cytochrome-c oxidase in using heme A as its active-site chromophores but lacks the CuA center and the cytochrome-c oxidizing activity of the mitochondrial enzyme. W...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00156a027
更新日期：1992-10-20 00:00:00
abstract：:It has been suggested that dimeric tubulin can participate in the signal transduction process through its association with the GTP-binding (G) proteins Gs and Gi1. Using the photoaffinity GTP analog, azidoanilido-GTP, it has been shown that the transfer of nucleotide from tubulin to G alpha s and G alpha i1 is the key...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00198a052
更新日期：1994-08-16 00:00:00
abstract：:The solution structure of the complementary d(C1-A2-T3-G4-X5-G6-T7-A8-C9).d(G10-T11-A12-C13-A14-C15-A16-T17-G18) DNA duplex (designated X.A 9-mer), which contains a 1,N2-propanodeoxyguanosine exocyclic adduct X5 opposite deoxyadenosine A14 at the center, is pH dependent [Kouchakdjian, M., Eisenberg, M., Live, D., Mari...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00066a004
更新日期：1993-04-20 00:00:00
abstract：:Peptide deformylase (EC 3.5.1.31) catalyzes the removal of a formyl group from the N-termini of nascent ribosome-synthesized polypeptides, an obligatory step during protein maturation in eubacteria. Since its discovery in crude Escherichia coli extracts 3 decades ago, the deformylase has resisted all attempts of purif...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi971155v
更新日期：1997-11-11 00:00:00
abstract：:The paramagnetic enhancements in the NMR relaxation rates for the fluorine in fluorophthalates have been used to determine the position of the phthalate with respect to the mononuclear metal ion in native and metal-substituted derivatives of phthalate dioxygenase (PDO). These studies show directly that the substrate i...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi990431y
更新日期：1999-08-24 00:00:00
abstract：:Fluid-phase interactions between hematologic cells and those of the vessel wall were studied in order to define a role for lipoxygenase products as cell signals in the control of vascular cholesterol metabolism. A functional parameter for hydroxy acids in this system has not been previously demonstrated. We report her...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00448a030
更新日期：1989-10-31 00:00:00
abstract：:PsbO, the photosystem II manganese stabilizing protein, contains an aspartate residue [Asp157 (spinach numbering)], which is highly conserved in eukaryotic and prokaryotic PsbOs. The homology model of the PSII-bound conformation of spinach PsbO presented here positions Asp157 in the large flexible loop of the protein....
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi9016999
更新日期：2009-12-22 00:00:00
abstract：:A critical point of interaction between F(1) and F(0) in the bacterial F(1)F(0)-ATP synthase is formed by the alpha and delta subunits. Previous work has shown that the N-terminal domain (residues 3-105) of the delta subunit forms a 6 alpha-helix bundle [Wilkens, S., Dunn, S. D., Chandler, J., Dahlquist, F. W., and Ca...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi0510678
更新日期：2005-09-06 00:00:00
abstract：:In order to further characterize the binding of the hybrid molecule NetOPC [bis(pyrrolecarboxamide)-oxazolopyridocarbazole conjugate] to double-stranded DNA, we have performed a molecular modeling study to investigate the binding modes of the complexes possibly formed between NetOPC and synthetic polynucleotides poly ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00172a018
更新日期：1994-02-15 00:00:00
abstract：:In addition to its superoxide dismutase (SOD) activity, Cu,Zn-superoxide dismutase (CuZnSOD) catalyzes the reductive decomposition of S-nitroso-L-glutathione (GSNO) in the presence of thiols such as L-glutathione (GSH). The GSNO-reductase activity but not the superoxide dismutase (SOD) activity of CuZnSOD is inhibited...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi061077o
更新日期：2006-10-24 00:00:00
abstract：:Coumermycin A(1) is a member of the aminocoumarin family of antibiotics. Unlike its structural relatives, novobiocin and clorobiocin, coumermycin A(1) is a dimer built on a 3-methyl-2,4-dicarboxypyrrole scaffold and bears two decorated noviose sugar components which are the putative target binding motifs for DNA gyras...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi048457z
更新日期：2004-11-30 00:00:00
abstract：:Previous experiments have shown that a Synechocystis sp. PCC 6803 mutant (delta psbO) lacking the extrinsic manganese-stabilizing protein (MSP) exhibits impaired, but significant levels of H2O-splitting activity [Burnap, R., & Sherman, L.A. (1991) Biochemistry 30, 440-446]. [14C]DCMU-binding experiments now show that ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00147a027
更新日期：1992-08-18 00:00:00
abstract：:The reverse transcriptase (RT) from the human immunodeficiency virus (HIV) is initially expressed as a 66-kDa protein and is subsequently proteolytically processed in vivo to form a 66-kDa/51-kDa heterodimer. Comparison of circular dichroism spectra of the 66-kDa, 51-kDa, and heterodimeric forms of RT indicates that t...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00150a015
更新日期：1992-09-08 00:00:00
abstract：:Isoform 1 of the mammalian Na(+)/H(+) exchanger (NHE1) is a ubiquitously expressed plasma membrane pH regulatory protein. It removes one intracellular H(+) in exchange for one extracellular Na(+). The 500 N-terminal amino acids comprise the catalytic membrane domain and fold into 12 transmembrane (TM) segments. To gai...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi500392y
更新日期：2014-06-10 00:00:00
abstract：:The Plasmodium falciparum malarial parasite genome appears to encode one and only one phosphatidylinositol 3'-kinase (PI3K), and sequence analysis suggests that the enzyme is a "class III"- or "Vps34"-type PI3K. PfVps34 has generated excitement as a possible druggable target and potentially a key target of artemisinin...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/acs.biochem.7b00416
更新日期：2017-08-22 00:00:00
abstract：:The present study uses CO as a surrogate for oxygen to probe how substrate binding triggers oxygen activation in peptidylglycine monooygenase (PHM). Infrared stretching frequencies (ν(C ≡ O)) of the carbonyl (CO) adducts of copper proteins are sensitive markers of Cu(I) coordination and are useful in probing oxygen re...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/acs.biochem.6b00845
更新日期：2016-12-06 00:00:00
abstract：:Catalase HPII from Escherichia coli is a homotetramer of 753 residue subunits. The multimer displays a number of unusual structural features, including interwoven subunits and a covalent bond between Tyr415 and His392, that would contribute to its rigidity and stability. As the temperature of a solution of HPII in 50 ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi982863z
更新日期：1999-03-30 00:00:00
abstract：:Human liver alcohol dehydrogenase [alcohol:NAD+ oxidoreductase, EC 1.1.1.1 (ADH)] catalyzes the stereospecific oxidation of different 3 beta-hydroxy-5 beta-steroids with ranges of Km from 46 to 320 microM and values of kcat from 7.0 to 72 min-1, pH 8.5. Only the class I isozymes containing gamma-subunits, gamma 1 gamm...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00412a013
更新日期：1988-06-14 00:00:00
abstract：:The local environment of the transmembrane and C-terminal domain of M13 major coat protein was probed by site-directed ESR spin labeling when the protein was introduced into three membrane-mimicking systems, DOPC vesicles, sodium cholate micelles, and SDS micelles. For this purpose, we have inserted unique cysteine re...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi961770j
更新日期：1996-12-03 00:00:00
abstract：:Coiled coils (CCs) are among the best-understood protein folds. Nonetheless, there are gaps in our knowledge of CCs. Notably, CCs are likely to be structurally more dynamic than often considered. Here, we explore this in an abundant class of CCs, parallel dimers, focusing on polar asparagine (Asn) residues in the hydr...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/acs.biochem.7b00848
更新日期：2017-12-19 00:00:00
abstract：:A method has been developed for calculating rate constants for dehydration of aldehydes that induce ATPase reactions by kinases and where 18O is transferred from the aldehyde or its hydrate to inorganic phosphate during the reaction. The method involves measurement of the fraction of 18O in phosphate by 31P NMR after ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00317a011
更新日期：1984-10-23 00:00:00
abstract：:Chicken liver phosphoenolpyruvate carboxykinase (PEPCK) is activated by Cr2+ as the sole activator under anaerobic conditions. PEPCK was modified with Cr3+, starting with either Cr2+ or Cr3+. Cr3+ has the distinct advantage of being a paramagnetic cation that could serve as a paramagnetic probe. Activators Mn2+, Mg2+,...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi9807299
更新日期：1998-06-02 00:00:00
abstract：:The effect of ionic strength on enzyme activity and stability varies considerably between enzymes. Ionic strength is known to affect the catalytic activity of some alkaline phosphatases (APs), such as Escherichia coli AP, but how ions affect APs is debated. Here, we studied the effect of various ions on a cold-adapted...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/acs.biochem.7b00690
更新日期：2017-09-26 00:00:00
abstract：:Thermal unfolding of discoidal complexes of apolipoprotein (apo) C-1 with dimyristoyl phosphatidylcholine (DMPC) reveals a novel mechanism of lipoprotein stabilization that is based on kinetics rather than thermodynamics. Far-UV CD melting curves recorded at several heating/cooling rates from 0.047 to 1.34 K/min show ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi025588w
更新日期：2002-06-11 00:00:00
abstract：:Heparin potentiates the mitogenic activity of FGF-1 by increasing the affinity for its receptor and by extending its biological half-life. During the course of labeling human FGF-1 with Na(125)I and chloramine T, it was observed that the protein lost its ability to bind to heparin. In contrast, bovine FGF-1 retained i...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi9903345
更新日期：1999-07-20 00:00:00
abstract：:The role of histidine in catalysis by prostaglandin H synthase has been investigated using chemical modification with diethyl pyrocarbonate (DEPC), an agent that has been found to rather selectively derivatize histidine residues in proteins under mild conditions. Incubation of the synthase apoprotein with DEPC at pH 7...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00124a013
更新日期：1992-03-10 00:00:00
abstract：:We investigated the molecular mechanisms and the binding site location for the fluorophor crystal violet (CrV), a noncompetitive antagonist of the nicotinic acetylcholine receptor (AChR). To this end, radiolabeled competition binding, fluorescence spectroscopy, Schild-type analysis, patch-clamp recordings, and molecul...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi051752e
更新日期：2006-02-21 00:00:00
