Abstract:
:All phosphagen kinases contain a conserved cysteine residue which has been shown by crystallographic studies, on both creatine kinase and arginine kinase, to be located in the active site. There are conflicting reports as to whether this cysteine is essential for catalysis. In this study we have used site-directed mutagenesis to replace Cys282 of human muscle creatine kinase with serine and methionine. In addition, we have replaced Cys282, conserved across all creatine kinases, with alanine. No activity was found with the C282M mutant. The C282S mutant showed significant, albeit greatly reduced, activity in both the forward (creatine phosphorylation) and reverse (MgADP phosphorylation) reactions. The K(m) for creatine was increased approximately 10-fold, but the K(m) for phosphocreatine was relatively unaffected. The V and V/K pH-profiles for the wild-type enzyme were similar to those reported for rabbit muscle creatine kinase, the most widely studied creatine kinase isozyme. However, the V/K(creatine) profile for the C282S mutant was missing a pK of 5.4. This suggests that Cys282 exists as the thiolate anion, and is necessary for the optimal binding of creatine. The low pK of Cys282 was also determined spectrophotometrically and found to be 5.6 +/- 0.1. The S284A mutant was found to have reduced catalytic activity, as well as a 15-fold increase in K(m) for creatine. The pK(a) of Cys282 in this mutant was found to be 6.7 +/- 0.1, indicating that H-bonding to Ser284 is an important, but not the sole, factor contributing to the unusually low pK(a) of Cys282.
journal_name
Biochemistryjournal_title
Biochemistryauthors
Wang PF,McLeish MJ,Kneen MM,Lee G,Kenyon GLdoi
10.1021/bi011208fsubject
Has Abstractpub_date
2001-10-02 00:00:00pages
11698-705issue
39eissn
0006-2960issn
1520-4995pii
bi011208fjournal_volume
40pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract::The search for a rational method of developing an antimalarial vaccine (malaria caused by Plasmodium falciparum) consists of blocking receptor-ligand interaction. Conserved peptides derived from proteins involved in invasion and having strong red blood cell binding ability have thus been identified; immunization studi...
journal_title:Biochemistry
pub_type: 历史文章,杂志文章
doi:10.1021/bi050214z
更新日期:2005-05-10 00:00:00
abstract::The temperature dependence of the reduction potential of the CuA site in carbon monoxide inhibited cytochrome c oxidase has been measured with a spectroelectrochemical method adapted to the relatively weak near-infrared absorption of this copper ion. These measurements, together with parallel measurements on the 604-n...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00349a024
更新日期:1986-01-14 00:00:00
abstract::A molecular model has been built of the equine infectious anemia virus (EIAV) proteinase on the basis of the crystal structures of the related Rous sarcoma virus (RSV) and human immunodeficiency virus (HIV) proteinases. The 104 residue long EIAV proteinase has 30 identical and 11 similar amino acids compared to those ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00064a019
更新日期:1993-04-06 00:00:00
abstract::Oxidative damage by transition metals bound to proteins may be an important pathogenic mechanism. Ceruloplasmin (Cp) is a Cu-containing plasma protein thought to be involved in oxidative modification of lipoproteins. We have previously shown that Cp increased cell-mediated low-density lipoprotein (LDL) oxidation by a ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi981137t
更新日期:1998-10-06 00:00:00
abstract::Surface plasmon resonance (SPR) spectroscopy has been used to follow incorporation and light-induced conformational changes in bovine rhodopsin reconstituted into an egg phosphatidylcholine bilayer deposited on a thin silver film. The magnitude of the SPR spectral changes caused by light varies with pH in a manner par...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00250a022
更新日期:1994-11-22 00:00:00
abstract::The virally encoded integrase protein carries out retroviral integration, which requires specific interactions with the two ends of the viral DNA, and also with host DNA that is the target of integration. We attached a photo-cross-linking agent to specific viral and target DNA sites to identify regions of the integras...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi972949c
更新日期:1998-05-12 00:00:00
abstract::A minor fraction of plasma high-density lipoprotein (pre beta-1 HDL) has been shown to promote cholesterol efflux from peripheral cell membranes [Castro, G. R., & Fielding, C. J. (1988) Biochemistry 27, 25-29]. When isolated native plasma is incubated at 37 degrees C, this fraction is specifically decreased. On the ot...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00160a022
更新日期:1992-11-17 00:00:00
abstract::Thermobifida fusca Cel9A-90, an unusual family 9 enzyme, is a processive endoglucanase containing a catalytic domain closely linked to a family 3c cellulose binding domain (Cel9A-68) followed by a fibronectin III-like domain and a family 2 cellulose binding domain. To study its catalytic mechanism, 12 mutant genes wit...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi049394n
更新日期:2004-08-03 00:00:00
abstract::The antiviral and antiproliferative activities of type I interferons (IFNs) are mediated by a common receptor, and its second subunit (IFNAR2) exhibits nanomolar affinity to both IFNalpha and IFNbeta subtypes. We have previously determined the structure of the IFN-binding extracellular domain of IFNAR2 (IFNAR2-EC) usi...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi049606g
更新日期:2004-08-10 00:00:00
abstract::Group I intron RNAs contain a core of highly conserved helices flanked by peripheral domains that stabilize the core structure. In the Tetrahymena group I ribozyme, the P4, P5, and P6 helices of the core pack tightly against a three-helix subdomain called P5abc. Chemical footprinting and the crystal structure of the T...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi982113p
更新日期:1999-03-09 00:00:00
abstract::Taq and T7 DNA polymerases have become basic molecular biology "tools" for DNA sequence analysis. However, Taq, unlike T7 DNA polymerase, is strongly biased against the incorporation of 2',3'-dideoxynucleotide triphosphates (ddNTPs) indicating very different substrate selectivities. Equilibrium binding and rate consta...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi951682j
更新日期:1996-02-20 00:00:00
abstract::Macromolecular protease inhibitors and camelid single-domain antibodies achieve their enzymic inhibition functions often through protruded structures that directly interact with catalytic centers of targeted proteases. Inspired by this phenomenon, we constructed synthetic human antibody libraries encoding long CDR-H3s...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.0c00690
更新日期:2020-10-13 00:00:00
abstract::Lateral mobilities of fluorescent cell surface probes have been measured on normal (3T3) and transformed (SV3T3) cultured mouse fibroblasts. There is little discernible difference in the mobilities of a lipid analogue (diI), a fluorescent ganglioside derivative (GM1), and tetramethylrhodamine-labeled succinylated conc...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00551a011
更新日期:1980-05-13 00:00:00
abstract::For the elucidation of the mechanism of steroid hormone receptor activation, the hydrodynamic properties of the unactivated and activated forms of the nonproteolyzed glucocorticoid receptor from the mouse AtT-20 pituitary tumor cell line were determined. The unactivated, molybdate-stabilized receptor has the following...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00277a038
更新日期:1983-04-12 00:00:00
abstract::The assignment of the 1H, 15N, 13CO, and 13C resonances of recombinant human interleukin-4 (IL-4), a protein of 133 residues and molecular mass of 15.4 kDa, is presented based on a series of 11 three-dimensional (3D) double- and triple-resonance heteronuclear NMR experiments. These studies employ uniformly labeled 15N...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00132a026
更新日期:1992-05-05 00:00:00
abstract::In recent years, a number of bioorthogonal reactions have been developed, exemplified by click chemistry, that enable the efficient formation of a specific product, even within a highly complex chemical environment. While the exquisite selectivity and reliability of these transformations have led to their broad applic...
journal_title:Biochemistry
pub_type: 杂志文章,评审
doi:10.1021/bi9007726
更新日期:2009-07-21 00:00:00
abstract::Calpain-catalyzed proteolysis of II-spectrin is a regulated event associated with neuronal long-term potentiation, platelet and leukocyte activation, and other processes. Calpain proteolysis is also linked to apoptotic and nonapoptotic cell death following excessive glutamate exposure, hypoxia, HIV-gp120/160 exposure,...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi061504y
更新日期:2007-01-16 00:00:00
abstract::Transmembrane chemoreceptors, also known as methyl-accepting chemotaxis proteins (MCPs), translate extracellular signals into intracellular responses in the bacterial chemotaxis system. MCP ligand binding domains control the activity of the CheA kinase, situated approximately 200 A away, across the cytoplasmic membran...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi801727m
更新日期:2009-03-10 00:00:00
abstract::A synthetic analogue of the tripeptide hemiasterlin, designated HTI-286, depolymerizes microtubules, is a poor substrate for P-glycoprotein, and inhibits the growth of paclitaxel-resistant tumors in xenograft models. Two radiolabeled photoaffinity analogues of HTI-286, designated 4-benzoyl-N,beta,beta-trimethyl-l-phen...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0474766
更新日期:2005-05-10 00:00:00
abstract::An attractive strategy for the development of anti-retroviral drugs is the exploration of compounds that mimic RNA control regions of the viral genome and act as "decoys" to sequester viral gene regulatory proteins. Decoys consisting of RNA, however, are chemically unstable and readily degraded by cellular nucleases. ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi961980w
更新日期:1997-01-21 00:00:00
abstract::The hypothesis that the stable ternary complex formed between myosin subfragment-1, MgADP and beryllium fluoride (BeF3-), denoted S-1 not equal to .ADP.BeF3-, is an analog of the intermediate state S-1**.ADP.P(i) has been tested in this work by examining the interactions of S-1 not equal to .ADP.BeF3- with actin. Equi...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00081a016
更新日期:1993-08-03 00:00:00
abstract::In the commonly used nucleation-dependent model of protein aggregation, aggregation proceeds only after a lag phase in which the concentration of energetically unfavorable nuclei reaches a critical value. The formation of oligomeric species prior to aggregation can be difficult to detect by current spectroscopic techn...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi400027y
更新日期:2013-03-19 00:00:00
abstract::Amyloid beta peptide (A beta) is released into the media of a variety of cells in culture during normal metabolism. The discovery of several missense mutations within or flanking the A beta region of the beta amyloid precursor protein (beta APP) in familial Alzheimer's disease provides strong evidence for a role of al...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00181a016
更新日期:1994-04-19 00:00:00
abstract::Serum paraoxonase hydrolyzes the toxic metabolites of a variety of organophosphorus insecticides. High serum paraoxonase levels appear to protect against the neurotoxic effects of organophosphorus substrates of this enzyme [Costa et al. (1990) Toxicol. Appl. Pharmacol. 103, 66-76]. The amino acid sequence accounting f...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00106a010
更新日期:1991-10-22 00:00:00
abstract::Studies on the structure-function relationship of transporters require the availability of sufficient amounts of the protein in a functional state. In this paper, we report the functional expression, purification, and reconstitution of the human sodium/d-glucose cotransporter1 (hSGLT1) in Pichia pastoris and ligand-in...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi051377q
更新日期:2005-11-29 00:00:00
abstract::The thermotropic behavior of aqueous dispersions of stearoylsphingomyelin-cholesterol mixtures was examined by high-sensitivity differential scanning calorimetry. When less than 20 mol % cholesterol was mixed with the sphingomyelin and the samples were held at room temperature for 7-9 days before the initiation of cal...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00528a010
更新日期:1981-12-08 00:00:00
abstract::Ca(2+)/calmodulin-dependent protein kinase kinases (CaMKKs) phosphorylate and activate specific downstream protein kinases, including CaMKI, CaMKIV, and 5'-AMP-activated protein kinase, which mediates a variety of Ca(2+) signaling cascades. CaMKKs have been shown to undergo autophosphorylation, although their role in ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi201005g
更新日期:2011-09-27 00:00:00
abstract::Overall activation volumes for multistep reactions are not usually pressure independent. The present investigation gives a quantitative description of this effect under Theory. Simple relations are obtained which can easily be applied to experimental data and which allow more insight into the dynamics of enzyme reacti...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00405a024
更新日期:1988-03-08 00:00:00
abstract::The ferritin IRE, a highly conserved (96-99% in vertebrates) mRNA translation regulatory element in animal mRNA, was studied by molecular modeling (using MC-SYM and DOCKING) and by NMR spectroscopy. Cobalt(III) hexammine was used to model hydrated Mg2+. IRE isoforms in other mRNAs regulate mRNA translation or stabilit...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9719814
更新日期:1998-02-10 00:00:00
abstract::Insulin and insulin-like growth factor 1 (IGF-1) share high sequence homology, but their folding behaviors are significantly different: insulin folds into one unique thermodynamically controlled structure, while IGF-1 folds into two thermodynamically controlled disulfide isomers. However, the origin of their different...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0346289
更新日期:2003-08-19 00:00:00