Abstract:
:Recombinant human 3'-phosphoadenosine 5'-phosphosulfate (PAPS) synthetase, isoform 1 (brain), was purified to near-homogeneity from an Escherichia coli expression system and kinetically characterized. The native enzyme, a dimer with each 71 kDa subunit containing an adenosine triphosphate (ATP) sulfurylase and an adenosine 5'-phosphosulfate (APS) kinase domain, catalyzes the overall formation of PAPS from ATP and inorganic sulfate. The protein is active as isolated, but activity is enhanced by treatment with dithiothreitol. APS kinase activity displayed the characteristic substrate inhibition by APS (K(I) of 47.9 microM at saturating MgATP). The maximum attainable activity of 0.12 micromol min(-1) (mg of protein)(-1) was observed at an APS concentration ([APS](opt)) of 15 microM. The theoretical K(m) for APS (at saturating MgATP) and the K(m) for MgATP (at [APS](opt)) were 4.2 microM and 0.14 mM, respectively. At likely cellular levels of MgATP (2.5 mM) and sulfate (0.4 mM), the overall endogenous rate of PAPS formation under optimum assay conditions was 0.09 micromol min(-1) (mg of protein)(-1). Upon addition of pure Penicillium chrysogenum APS kinase in excess, the overall rate increased to 0.47 micromol min(-1) (mg of protein)(-1). The kinetic constants of the ATP sulfurylase domain were as follows: V(max,f) = 0.77 micromol min(-1) (mg of protein)(-1), K(mA(MgATP)) = 0.15 mM, K(ia(MgATP)) = 1 mM, K(mB(sulfate)) = 0.16 mM, V(max,r) = 18.7 micromol min(-1) (mg of protein)(-1), K(mQ(APS)) = 4.8 microM, K(iq(APS)) = 18 nM, and K(mP(PPi)) = 34.6 microM. The (a) imbalance between ATP sulfurylase and APS kinase activities, (b) accumulation of APS in solution during the overall reaction, (c) rate acceleration provided by exogenous APS kinase, and (d) availability of both active sites to exogenous APS all argue against APS channeling. Molybdate, selenate, chromate ("chromium VI"), arsenate, tungstate, chlorate, and perchlorate bind to the ATP sulfurylase domain, with the first five serving as alternative substrates that promote the decomposition of ATP to AMP and PP(i). Selenate, chromate, and arsenate produce transient APX intermediates that are sufficiently long-lived to be captured and 3'-phosphorylated by APS kinase. (The putative PAPX products decompose to adenosine 3',5'-diphosphate and the original oxyanion.) Chlorate and perchlorate form dead-end E.MgATP.oxyanion complexes. Phenylalanine, reported to be an inhibitor of brain ATP sulfurylase, was without effect on PAPS synthetase isoform 1.
journal_name
Biochemistryjournal_title
Biochemistryauthors
Lansdon EB,Fisher AJ,Segel IHdoi
10.1021/bi049827msubject
Has Abstractpub_date
2004-04-13 00:00:00pages
4356-65issue
14eissn
0006-2960issn
1520-4995journal_volume
43pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract::In eukaryotes, a major route for ATP-dependent protein breakdown proceeds through covalent intermediates of target proteins destined for degradation and the highly conserved, 76 amino acid protein ubiquitin. In rabbit reticulocytes, it has been shown that hemin effectively inhibits this pathway by blocking the catabol...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00409a025
更新日期:1988-05-03 00:00:00
abstract::Electron paramagnetic resonance (EPR) measurements were performed on photosystem II (PSII) membranes that were treated with 2 M NaCl to release the 17- and 23-kilodalton (kDa) polypeptides. By using 75 microM 3-(3,4-dichlorophenyl)-1,1-dimethylurea to limit the photosystem II samples to one stable charge separation in...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00369a022
更新日期:1986-10-21 00:00:00
abstract::Although the importance of interactions involving both the cytosolic and transmembrane regions of SecY and SecE has been documented, no information has been available for the physical contact sites of these translocase subunits in their cytosolic domains. We now carried out site-specific cross-linking experiments to i...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi034333v
更新日期:2003-06-24 00:00:00
abstract::Tryptophan 2,3-dioxygenase (TDO) from Xanthomonas campestris is a highly specific heme-containing enzyme from a small family of homologous enzymes, which includes indoleamine 2,3-dioxygenase (IDO). The structure of wild type (WT TDO) in the catalytically active, ferrous (Fe (2+)) form and in complex with its substrate...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi801202a
更新日期:2008-10-07 00:00:00
abstract::Stimulation of isolated hepatocytes with epidermal growth factor (EGF) causes rapid tyrosine phosphorylation of the EGF receptor (EGFR) and adapter/target proteins, which was monitored with 1 and 2 s resolution at 37, 20, and 4 degrees C. The temporal responses detected for multiple signaling proteins involve both tra...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi011506c
更新日期:2002-01-08 00:00:00
abstract::The biochemical and structural properties of bovine retinal nucleoside diphosphate kinase were investigated. The enzyme showed two polypeptides of approximately 17.5 and 18.5 kDa on SDS-PAGE, while isoelectric focusing revealed seven to eight proteins with a pI range of 7.4-8.2. Sedimentation equilibrium yielded a mol...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi980853s
更新日期:1998-10-06 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::Human granulocyte chemotactic protein 2 (GCP-2) has originally been isolated from cytokine-stimulated osteosarcoma cells as a chemokine coproduced in minute amounts together with interleukin 8. Human GCP-2 (75 residues) was synthesized on a 0.25-mmol scale using Fmoc chemistry. After disulfide bridge formation and pur...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi961999z
更新日期:1997-03-04 00:00:00
abstract::Adenosine 5'-phosphosulfate (APS) kinase catalyzes the second reaction in the two-step conversion of inorganic sulfate to 3'-phosphoadenosine 5'-phosphosulfate (PAPS). This report presents the 2.0 A resolution crystal structure of ligand-free APS kinase from the filamentous fungus, Penicillium chrysogenum. The enzyme ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9924157
更新日期:2000-02-22 00:00:00
abstract::A small metal-binding protein (SmbP) with no known similarity to other proteins in current databases was isolated and characterized from the periplasm of Nitrosomonas europaea. The primary structure of this small (9.9 kDa) monomeric protein is characterized by a series of 10 repeats of a seven amino acid motif and an ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi049318k
更新日期:2004-09-07 00:00:00
abstract::Phospholipase B1 (PLB1), secreted by the pathogenic yeast Cryptococcus neoformans, has an established role in virulence. Although the mechanism of its phospholipase B, lysophospholipase, and lysophospholipase transacylase activities is unknown, it possesses lipase, subtilisin protease aspartate, and phospholipase moti...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi7009508
更新日期:2007-09-04 00:00:00
abstract::The Escherichia coli L-arabinose-binding protein (ABP) participates as a specific receptor in the transport of L-arabinose, D-fucose, and D-galactose through the periplasmic space. The wild-type protein binds L-arabinose about 40 times more strongly than D-fucose. A mutation of the protein at position 108 (Met-->Leu) ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00080a013
更新日期:1993-07-27 00:00:00
abstract::We have used electron spin resonance and circular dichroism to examine and compare the dynamics in two analogues of the Ala-based 3K(I) peptide [Marqusee, S., Robbins, V.H., & Baldwin, R. L. (1989) Proc. Natl. Acad. Sci. U.S.A. 86, 5286-5290], labeled at positions 4 and 8, throughout the alpha-helix----coil transition...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00103a016
更新日期:1991-10-01 00:00:00
abstract::Fom3, the antepenultimate enzyme in the fosfomycin biosynthetic pathway in Streptomyces spp., is a class B cobalamin-dependent radical SAM methyltransferase that catalyzes methylation of (5'-cytidylyl)-2-hydroxyethylphosphonate (2-HEP-CMP) to form (5'-cytidylyl)-2-hydroxypropylphosphonate (2-HPP-CMP). Previously, the ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.8b00616
更新日期:2018-08-21 00:00:00
abstract::The interactions between rhodopsin molecules in a micellar detergent solution (octyl glucoside) and in reconstituted phospholipid vesicles were studied in the dark and after bleaching. Resonance energy transfer measurements were used to monitor the proximity between rhodopsin monomers conjugated with a fluorescent don...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00270a030
更新日期:1983-01-04 00:00:00
abstract::Since one of the electrophysiological effects of pyrithiamin, an antimetabolite of thiamin, suggested an interference with sodium pump mechanisms, the effect of pyrithiamin on Na+,K+-ATPase was investigated. We found that whereas preincubation of the antimetabolite with nonneuronal preparations of Na+,K+-ATPase produc...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00278a023
更新日期:1983-04-26 00:00:00
abstract::In synthetic biology, metabolic engineering, and gene therapy, there is a strong demand for orthogonal or externally controlled regulation of gene expression. Here, RNA-based regulatory devices represent a promising emerging alternative to proteins, allowing a fast and direct control of gene expression, as no synthesi...
journal_title:Biochemistry
pub_type: 杂志文章,评审
doi:10.1021/acs.biochem.6b01218
更新日期:2017-03-07 00:00:00
abstract::Photoactivated adenylyl cyclases are powerful tools for optogenetics and for investigating signal transduction mechanisms in biological photoreceptors. Because of its large increase in enzyme activity in the light, the BLUF (blue light sensor using flavin adenine dinucleotide)-activated adenylyl cyclase (bPAC) from Be...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi500479v
更新日期:2014-08-12 00:00:00
abstract::Baeyer-Villiger monooxygenases (BVMOs), mostly flavoproteins, were shown to be powerful biocatalysts for synthetic organic chemistry applications and were also suggested to play key roles for the biosyntheses of various natural products. Here we present the three-dimensional structure of MtmOIV, a 56 kDa homodimeric F...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi8023509
更新日期:2009-06-02 00:00:00
abstract::The isolated N-terminal Src homology 3 (SH3) domain of Drosophila drk exists in equilibrium between folded and unfolded states in aqueous buffer near neutral pH. Nuclear magnetic resonance spectra recorded on both states simultaneously exhibit an approximate 1:1 ratio of protein conformations. The folded form is simil...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00020a025
更新日期:1995-05-23 00:00:00
abstract::The complex kinetic behavior commonly observed in protein folding studies suggests that a heterogeneous population of molecules exists in solution and that a number of discrete steps are involved in the conversion of unfolded molecules to the fully native form. A central issue in protein folding is whether any of thes...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9700476
更新日期:1997-05-13 00:00:00
abstract::Elucidation of the molecular mechanism leading to biomembrane fusion is a challenging issue in current biomedical research in view of its involvement in controlling cellular functions and in mediating various important diseases. According to the generally admitted stalk mechanism described for membrane fusion, negativ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi060317x
更新日期:2006-08-01 00:00:00
abstract::Cells cultured from human skin synthesize precursor forms of types I and III collagens. After denaturation and reduction, the polypeptide chains obtained from these molecules had molecular weights estimated to be 140,000 and 120, 000. The larger chains, the pro alpha chains, are believed to be derived from the origina...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00679a007
更新日期:1975-04-22 00:00:00
abstract::Di- and triketone inhibitors of (4-hydroxyphenyl)pyruvate dioxygenase (HPPD) are both effective herbicides and therapeutics. The inhibitory activity is used to halt the production of lipophilic redox cofactors in plants and also in humans to prevent accumulation of toxic metabolic byproducts that arise from specific i...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi049317s
更新日期:2004-06-01 00:00:00
abstract::The azinomycins are a family of potent antitumor agents with the ability to form interstrand cross-links with DNA. This study reports on the unusual biosynthetic formation of the 5-methyl naphthoate moiety, which is essential for effective DNA association. While sequence analysis predicts that the polyketide synthase ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.5b01050
更新日期:2016-02-02 00:00:00
abstract::The gene encoding enzyme I of the phosphoenolpyruvate:sugar phosphotransferase system from an Escherichia coli enzyme I mutant was cloned and sequenced. The mutation was shown to be a guanine to adenine transition resulting in an altered protein in which glycine-338 was replaced by aspartic acid. The enzyme I structur...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi952052k
更新日期:1996-01-09 00:00:00
abstract::Transport of hydrophilic di- and tripeptides into Lactococcus lactis is mediated by a proton motive force-driven peptide transport protein (DtpT) that shares similarity with eukaryotic peptide transporters, e.g., from kidney and small intestine of rabbit, man, and rat. Hydropathy profiling in combination with the "pos...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi963068t
更新日期:1997-06-03 00:00:00
abstract::Globotriaosylceramide [(Gal alpha 1-4Gal beta 1-4Glc-ceramide (Gb3)] was separated from human kidney, and the fatty acid composition was determined. Semisynthetic Gb3 molecular species of corresponding fatty acid chain length were prepared and compared for verotoxin (VT) binding affinity by TLC overlay, and a quantita...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00120a011
更新日期:1992-02-11 00:00:00
abstract::In the reaction center (RC) of the photosynthetic bacterium Rhodobacter sphaeroides, two ubiquinone molecules, QA and QB, play a pivotal role in the conversion of light energy into chemical free energy by coupling electron transfer to proton uptake. In native RCs, the transfer of an electron from QA to QB takes place ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi700297b
更新日期:2007-04-17 00:00:00
abstract::Adenylosuccinate lyase of Bacillus subtilis is inactivated by 2-[(4-bromo-2,3-dioxobutyl)thio]adenosine 5'-monophosphate (2-BDB-TAMP) at pH 7.0. As the reagent concentration is increased, a maximum rate constant is approached, indicative of reversible enzyme-reagent complex formation (KR = 68 +/- 9 microM) prior to ir...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9805339
更新日期:1998-06-09 00:00:00