Abstract:
:CopA, a thermophilic membrane ATPase from Archaeoglobus fulgidus, drives the outward movement of Cu(+) or Ag(+) [Mandal et al. (2002) J. Biol. Chem. 277, 7201-7208]. This, as other P(IB)-ATPases, is characterized by a putative metal binding sequence (C(380)PC(382)) in its sixth transmembrane fragment and cytoplasmic metal binding sequences in its NH(2)- and COOH-terminal ends (C(27)AMC(30) and C(751)HHC(754)). Using isolated CopA, we have studied the functional role of these three putative metal binding domains. Replacement of transmembrane Cys residues by Ala results in nonfunctional enzymes that are unable to hydrolyze ATP. However, the CPC --> APA substituted enzyme binds ATP, indicating its correct folding and suggesting that enzyme turnover is prevented by the lack of metal binding to the transmembrane site. Replacement of C-terminal Cys by Ala (C(751,754)A) has no significant effect on ATPase activity, enzyme phosphorylation, apparent binding affinities of ligands, or E1-E2 equilibrium. In contrast, replacement of Cys in the N-terminal metal binding domain (N-MBD) (C(27,30)A) leads to 40% reduction in enzyme turnover. The C(27,30)A enzyme binds Cu(+), Ag(+), and ATP with the same high apparent affinities as the wild-type CopA. Evidence that N-MBD disruption has no effect on the E1-E2 equilibrium is provided by the normal interaction of ATP acting with low affinity and the unaffected IC(50) for vanadate inhibition observed in the C(27,30)A-substituted enzyme. However, replacement C(27,30)A slowed the dephosphorylation of the E2P(metal) form of the enzyme, suggesting a reduction in the rate of metal release. Other investigators have shown the Cu-dependent interaction of isolated N-MBDs from the Wilson disease Cu-ATPase with the ATP binding cytoplasmic domain [Tsivkovskii et al. (2001) J. Biol. Chem. 276, 2234-2242]. Therefore, the data suggest a regulatory mechanism in which the Cu-dependent N-MBD/ATP binding domain interaction would accelerate cation release, the enzyme rate-limiting step, and consequently Cu(+) transport.
journal_name
Biochemistryjournal_title
Biochemistryauthors
Mandal AK,Argüello JMdoi
10.1021/bi034806ysubject
Has Abstractpub_date
2003-09-23 00:00:00pages
11040-7issue
37eissn
0006-2960issn
1520-4995journal_volume
42pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract::Eukaryotic transfer RNA precursors (pre-tRNAs) contain a 5' leader preceding the aminoacyl acceptor stem and a 3' trailer extending beyond this stem. An early step in pre-tRNA maturation is removal of the 5' leader by the endoribonuclease, RNase P. Extensive pairing between leader and trailer sequences has previously ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi000603n
更新日期:2000-08-15 00:00:00
abstract::The ability of a class of C-20' modified vinca alkaloid congeners to induce tubulin spiral formation was investigated relative to their ability to inhibit microtubule assembly, their cytotoxicity against a leukemic cell line, L1210, and their measured and calculated partition coefficients. These studies were prompted ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi001038r
更新日期:2000-10-03 00:00:00
abstract::PI-SceI is a member of an unusual class of rare cutting homing endonucleases produced by an autocatalytic protein splicing from a precursor. To analyze the structural and functional domain organization of the endonuclease PI-SceI and to examine whether the DNA binding activity can be structurally separated from the ca...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi980013d
更新日期:1998-06-02 00:00:00
abstract::A procedure was developed for selective radiolabeling of membrane proteins on cells to higher specific activities than possible with available techniques. Cell surface amino groups were derivatized with 125I-(hydroxyphenyl)propionyl groups via 125I-sulfosuccinimidyl (hydroxyphenyl)propionate (125I-sulfo-SHPP). This re...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00377a014
更新日期:1987-02-10 00:00:00
abstract::Bean-deleterious Pseudomonas A214 produced the extracellular yellow-green, fluorescent siderophore [microbial iron(III) transport agent] pseudobactin A214 under iron-limiting conditions. Pseudobactin A214 has a molecular formula of C46H64N13O22 and a molecular mass of 1151 g/mol. Pseudobactin A214 contained an N-block...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00367a022
更新日期:1986-09-23 00:00:00
abstract::Protease inhibition by metastable serine protease inhibitors (serpins) is mediated by one of the largest functional intradomain conformational changes known in biology. In this extensive structural rearrangement, protease-serpin complex formation triggers cleavage of the serpin reactive center loop (RCL), its subseque...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/acs.biochem.6b00839
更新日期:2017-01-31 00:00:00
abstract::The X-ray crystal structure of a salicylhydroxamic acid (SHA) inhibitory complex with human myeloperoxidase (MPO) has been determined at 2.3 A resolution. The aromatic ring of the inhibitor binds to a hydrophobic region at the entrance to the distal heme pocket between heme pyrrole ring D and the side chain of Arg 239...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi960577m
更新日期:1996-08-20 00:00:00
abstract::Malic enzyme from pigeon liver is alkylated by the substrate analogue bromopyruvate, resulting in the concomitant loss of its oxidative decarboxylase and oxalacetate decarboxylase activities, but not its ability to reduce alpha-keto acids. The inactivation of oxidative decarboxylase activity follows saturation kinetic...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00621a024
更新日期:1977-01-25 00:00:00
abstract::Endonuclease V of bacteriophage T4 binds to UV-irradiated deoxyribonucleic acid (DNA) but not to unirradiated DNA. We have developed an assay to detect this binding, based on the retention of enzyme--DNA complexes on nitrocellulose filters. The amount of complex retained, ascertained by using radioactive DNA, is a mea...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00549a026
更新日期:1980-04-15 00:00:00
abstract::UDP-GlcNAc 2-epimerase/ManNAc kinase (GNE) catalyzes the first two committed steps in sialic acid synthesis. In addition to the three previously described human GNE isoforms (hGNE1-hGNE3), our database and polymerase chain reaction analysis yielded five additional human isoforms (hGNE4-hGNE8). hGNE1 is the ubiquitousl...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi201050u
更新日期:2011-10-18 00:00:00
abstract::Heparin was isolated from an unusually large human hemangioma that contained an elevated level of mast cells. Purification of multimilligram quantities of heparin from this tissue sample permitted a thorough examination of its structure and activity. Characterization of this human heparin included the following: one-d...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00164a020
更新日期:1992-12-15 00:00:00
abstract::The heat production for the reaction steps of ATP hydrolysis by heavy meromyosin has been reexamined by using a reaction-type calorimeter. So far, the values reported for the heat production have varied substantially among research groups. The present results obtained in 0.3 M KCl, 10 mM MgCl2, and 20 mM Tris-HCl at p...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00518a037
更新日期:1981-07-21 00:00:00
abstract::The p90 ribosomal S6 family of kinases (RSK) are potential drug targets, due to their involvement in cancer and other pathologies. There are currently only two known selective inhibitors of RSK, but the basis for selectivity is not known. One of these inhibitors is a naturally occurring kaempferol-α-L-diacetylrhamnosi...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi300620c
更新日期:2012-08-21 00:00:00
abstract::By limited proteolysis of a type kappa immunoglobulin light chain (Oku) with clostripain, both the VL and CL fragments were obtained with a high yield. The unfolding and refolding by guanidine hydrochloride of light chain Oku and its VL and CL fragments were studied at pH 7.5 and 25 degrees C with circular dichroism a...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00393a015
更新日期:1987-09-22 00:00:00
abstract::The Ca2+-ATPase of sarcoplasmic reticulum was purified and depleted of proteolipids by solubilization in Triton X-100 and by fractionation on a DE-52 column. The protein reconstituted by deoxycholate-cholate dialysis at low lipid to protein ratios (2-5 mg of lipid/mg of protein), with either dioleoylphosphatidylethano...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00296a021
更新日期:1984-01-03 00:00:00
abstract::Replacement of methionine (Met) residues by selenomethionine (SeMet) was recently shown to facilitate the crystallographic analysis of protein structure through the application of multi-wavelength anomalous diffraction techniques [Yang et al. (1990) Science (Washington, D.C.) 249, 1398-1405]. The availability of SeMet...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00110a009
更新日期:1991-11-19 00:00:00
abstract::Pyrophosphate-dependent 6-phosphofructo-1-kinase (PPi-PFK) from Propionibacterium freudenreichii was inactivated by low concentrations of the lysine-specific reagent pyridoxal phosphate (PLP) after sodium borohydride reduction. The substrates fructose 6-phosphate and fructose 1,6-bisphosphate protected against inactiv...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00135a011
更新日期:1992-05-26 00:00:00
abstract::Src SH3 is a small all-beta-sheet protein composed of a single domain. We studied the folding behavior of src SH3 at various conditions by circular dichroism (CD), fluorescence, and X-ray solution scattering methods. On the src SH3 folding pathway, an alpha-helix-rich intermediate appeared not only at subzero temperat...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi0618262
更新日期:2007-05-01 00:00:00
abstract::Monoamine oxidase B (MAO B) is an integral protein of the outer mitochondrial membrane that is involved in the deamination of vasoactive and neuroactive amines. The oxidation of these amine substrates requires the cofactor FAD, which is covalently bound to Cys-397 of human MAO B. Previously, Glu-34 and Tyr-44 of MAO B...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi9806910
更新日期:1998-09-01 00:00:00
abstract::Alpha-synuclein modulates dopamine homeostasis in dopamine-producing neurons of substantia nigra, partly through regulation of human dopamine transporter (hDAT) activity. To identify the underlying mechanisms, we disrupted the modulation of hDAT activity by wild-type (wt) alpha-synuclein, and its familial Parkinson's ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi035308s
更新日期:2004-02-10 00:00:00
abstract::Modification of the bacterial cell wall heteropolymer peptidoglycan by addition of an acetyl group to the C-6 hydroxyl group of N-acetylmuramoyl residues is known to inhibit the activity of muramidases (lysozymes) of innate immune systems. The O-acetylation of peptidoglycan also precludes the action of intrinsic lytic...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi051679s
更新日期:2006-01-24 00:00:00
abstract::As part of a larger effort to engineer the stability and hemin-binding properties of microsomal (Mc) cytochromes b(5) into rat liver outer mitochondrial membrane (OM) cytochrome (cyt) b(5), several mutants of rat OM cyt b(5) were prepared to study the effect of gradual and complete elimination of two extended hydropho...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi026005l
更新日期:2002-10-01 00:00:00
abstract::The newly discovered oncogenic protein gankyrin, which contains six ankyrin repeats, has been reported to be involved in the phosphorylation and degradation of the retinoblastoma gene product, Rb. Using in vitro systems, we have identified a peptide fragment of gankyrin, 176LHLACDEERN185, which is responsible for bind...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi011550s
更新日期:2002-03-26 00:00:00
abstract::The ubiquitin signaling pathway consists of hundreds of enzymes that are tightly regulated for the maintenance of cell homeostasis. Parkin is an E3 ubiquitin ligase responsible for conjugating ubiquitin onto a substrate protein, which itself can be ubiquitinated. Ataxin-3 performs the opposing function as a deubiquiti...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi400780v
更新日期:2013-10-22 00:00:00
abstract::The photoaddition of 4'-(hydroxymethyl)-4,5',8-trimethylpsoralen (HMT) to different conformational states of RNA was studied. Poly(U), poly(A,U) (random copolymer), poly(A-U) (alternating copolymer), poly(A) . poly(U) (double stranded), and poly(U) . poly(A) . poly(U) (triple stranded) were reacted with HMT at differe...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00535a040
更新日期:1982-03-16 00:00:00
abstract::Neutrophil elastase (NE), a mediator of inflammation, binds with high affinity numerous anionic molecules including suramin, a polysulfated naphthylurea, which inhibits it with a Ki of 0.2 microM and a 4:1 suramin:NE stoichiometry and thus constitutes a potential therapeutic agent. In an attempt to locate the suramin ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi971029r
更新日期:1997-12-16 00:00:00
abstract::The ligand-binding dynamics and the reaction with O(2) of the fully (five-electron) reduced cytochrome caa(3) from the thermohalophilic bacterium Rhodothermus (R.) marinus were investigated. The enzyme is a proton pump which has all the residues of the proton-transfer pathways found in the mitochondrial-like enzymes c...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi010344h
更新日期:2001-09-04 00:00:00
abstract::The Synechococcus PCC7942 SmtB is a zinc-responsive transcriptional repressor and a member of the ArsR superfamily of prokaryotic metalloregulatory transcription factors. The mechanism of negative regulation by Zn(II) and other metals as well as the coordination chemistry (stoichiometry, affinity, and specificity) of ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi001140o
更新日期:2000-09-26 00:00:00
abstract::The reactivity of aluminum and several other divalent and trivalent metallic cations toward chromatin from rat brain and liver has been investigated. Two criteria are used to determine the relative reactivity of these cations toward chromatin. The first involves the ability of the ions to compact the chromatin fibers ...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi00435a043
更新日期:1989-05-02 00:00:00
abstract::Primase DnaG is an essential bacterial enzyme that synthesizes short ribonucleotide primers required for chromosomal DNA replication. Inhibitors of DnaG can serve as leads for development of new antibacterials and biochemical probes. We recently developed a nonradioactive in vitro primase-pyrophosphatase assay to iden...
journal_title:Biochemistry
pub_type: 杂志文章
doi:10.1021/bi4011286
更新日期:2013-10-01 00:00:00