Abstract：
:Salmonella typhimurium L-histidinol dehydrogenase (EC 1.1.1.23), a four-electron dehydrogenase, was inactivated by an active-site-directed modification reagent, 7-chloro-4-nitro-2,1,3-benzoxadiazole (NBD-Cl). The inactivation followed pseudo-first-order kinetics and was prevented by low concentrations of the substrate L-histidinol or by the competitive inhibitors histamine and imidazole. The observed rate saturation kinetics for inactivation suggest that NBD-Cl binds to the enzyme noncovalently before covalent inactivation occurs. The UV spectrum of the inactivated enzyme showed a peak at 420 nm, indicative of sulfhydryl modification. Stoichiometry experiments indicated that full inactivation was correlated with modification of 1.5 sulfhydryl groups per subunit of enzyme. By use of a substrate protection scheme, it was shown that 0.5 sulfhydryl per enzyme subunit was neither protected against NBD-Cl modification by L-histidinol nor essential for activity. Modification of the additional 1.0 sulfhydryl caused complete loss of enzyme activity and was prevented by L-histidinol. Pepsin digestion of NBD-modified enzyme was used to prepare labeled peptides under conditions that prevented migration of the NBD group. HPLC purification of the peptides was monitored at 420 nm, which is highly selective for NBD-labeled cysteine residues. By amino acid sequencing of the major peptides, it was shown that the reagent modified primarily Cys-116 and Cys-377 and that the presence of L-histidinol gave significant protection of Cys-116. The presence of a cysteine residue in the histidinol binding site is consistent with models in which formation and subsequent oxidation of a thiohemiacetal occurs as an intermediate step in the overall reaction.
journal_name
Biochemistryjournal_title
Biochemistryauthors
Grubmeyer CT,Gray WRdoi
10.1021/bi00365a009subject
Has Abstractpub_date
1986-08-26 00:00:00pages
4778-84issue
17eissn
0006-2960issn
1520-4995journal_volume
25pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract：:These investigations characterize the covalent binding of reactive products of prostaglandin H-synthases (PGHSs) to the enzyme and to other molecules. The intermediate product of oxygenation of arachidonic acid by the PGHSs, prostaglandin (PG) H2, undergoes rearrangement to the highly reactive gamma-keto aldehydes, le...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi002629k
更新日期：2001-06-12 00:00:00
abstract：:The bacterial DNA cytosine methyltransferase M.HhaI sequence-specifically modifies DNA in an S-adenosylmethionine dependent reaction. The enzyme stabilizes the target cytosine (GCGC) into an extrahelical position, with a concomitant large movement of an active site loop involving residues 80-99. We used multidimension...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi602662e
更新日期：2007-06-19 00:00:00
abstract：:Despite significant progress in the past decade, questions still remain about the complete structural, dynamic, and thermodynamic effect of the cis-syn cyclobutane pyrimidine dimer lesion (hereafter called the thymine dimer) on double-stranded genomic DNA. We examined a 19-mer oligodeoxynucleotide duplex containing a ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi801417u
更新日期：2008-12-09 00:00:00
abstract：:The three-dimensional chalice-like crystal structure of initiation factor 2 IF2/eIF5B from Methanobacterium thermoautotrophicum represents a novel fold and domain architecture in which the N-terminal G domain and the C-terminal C domain are separated by an approximately 40 A alpha-helix. Homologous Thermus thermophilu...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi702295g
更新日期：2008-04-29 00:00:00
abstract：:The value of reducing the activity of ornithine decarboxylase (ODC), a key enzyme in the biosynthesis of polyamines, is well-appreciated. Polyamines are necessary components for cell growth, and manipulation of polyamine homeostasis may be an effective strategy for the treatment of a number of disorders, including neo...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi047918d
更新日期：2005-02-15 00:00:00
abstract：:The occupancy of the central cavity of the membrane-embedded c ring of the ATP synthase of Escherichia coli was investigated with a photo-cross-linking approach. Single cysteine mutants were created at c subunit positions 4, 8, and 11, which are oriented to the inside of the ring. These cysteines were alkylated with r...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi052304+
更新日期：2006-02-14 00:00:00
abstract：:The peptides generated from the degradation of the oxidized B chain of bovine insulin by the multiproteinase complex macropain (proteasome) have been analyzed by reverse-phase peptide mapping and identified by N-terminal amino acid sequencing and composition analysis. Six of the 29 peptide bonds in the insulin B chain...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00224a022
更新日期：1991-03-12 00:00:00
abstract：:Cytochrome P450 reductase (CPR) is a tethered membrane protein which transfers electrons from NADPH to microsomal P450s. We show that the lipid bilayer has a role in defining the redox potential of the CPR flavin domains. In order to quantitate the electrochemical behavior of this central redox protein, full-length CP...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi9011435
更新日期：2009-12-29 00:00:00
abstract：:Protein disulfide-isomerase, a protein localized to the lumen of the endoplasmic reticulum of eukaryotic cells, catalyzes the posttranslational formation and rearrangement of protein disulfide bonds. As isolated from bovine liver, the enzyme contains 0.8 free sulfhydryl group per mole of protein monomer and 3.1 disulf...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00444a023
更新日期：1989-09-05 00:00:00
abstract：:We have measured the microsecond rotational motions of myosin heads in contracting rabbit psoas muscle fibers by detecting the transient phosphorescence anisotropy of eosin-5-maleimide attached specifically to the myosin head. Experiments were performed on small bundles (10-20 fibers) of glycerinated rabbit psoas musc...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00495a003
更新日期：1990-10-30 00:00:00
abstract：:G-protein-coupled receptors (GPCRs) have evolved as highly specialized cellular machinery that can dictate biological outcomes in response to diverse stimuli. Specifically, they induce multiple pathway responses upon structural perturbations induced at local protein sites. GPCRs utilize a concurrent strategy involving...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/acs.biochem.9b00884
更新日期：2020-01-28 00:00:00
abstract：:The first step in sulfur mobilization for the biosynthesis of Fe-S clusters under oxidative stress and iron starvation in Escherichia coli involves a cysteine desulfurase SufS. Its catalytic reactivity is dependent on the presence of a sulfur acceptor protein, SufE, which acts as the preferred substrate for this enzym...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi4001479
更新日期：2013-06-11 00:00:00
abstract：:Hydroxyproline-rich glycoproteins (HRGPs) are the major proteinaceous components of higher plant walls and the predominant components of the cell wall of the green alga Chlamydomonas reinhardtii. The GP1 protein, an HRGP of the C. reinhardtii wall, is shown to adopt a polyproline II helical configuration and to carry ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi0023605
更新日期：2001-03-06 00:00:00
abstract：:Comparative electrophoretic, thermal denaturation, and spectroscopic studies with dodecamers of the form d(ATTA-XGCX-TAAT) and their self-complementary counterparts suggest that actinomycin D (ACTD) binds strongly to a 5'GC3' site with flanking T/T mismatches and moderately to that with C/C mismatches but weakly to th...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi961060d
更新日期：1996-12-17 00:00:00
abstract：:The isolated cytochrome (cyt) b(6)f complex from spinach is inhibited by Cu(2+) with a K(D) of about 1 microM at pH 7.6 in the presence of 1.6 microM decyl-plastoquinol (C(10)-PQH(2)) as a substrate. Inhibition was competitive with respect to C(10)-PQH(2) but noncompetitive with respect to horse heart cyt c or plastoc...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi991974a
更新日期：2000-03-28 00:00:00
abstract：:We found that recombinant human adult hemoglobin (rHb A) expressed in Escherichia coli showed heterogeneity of components with the intensity of a positive CD band at 260 nm and that it could be resolved into three components (SP-1, SP-2, and SP-3) by SP-Sepharose column chromatography. 1H NMR revealed that SP-1 is ide...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi7015519
更新日期：2008-01-15 00:00:00
abstract：:Soluble guanylate cyclase (sGC) is a heterodimeric hemoprotein composed of alpha1 and beta1 subunits. sGC is activated by nitric oxide (NO) and therefore plays a central role in NO signal transduction. Activation of sGC by NO is believed to be mediated by the interaction between NO and the heme of sGC. Spectroscopic a...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi971825x
更新日期：1997-12-16 00:00:00
abstract：:The protein phosphatase 2C (PP2C) family represents one of the four major protein Ser/Thr phosphatase activities in mammalian cells and contains at least 13 distinct gene products. Although PP2C family members regulate a variety of cellular functions, mechanisms of regulation of their activities are largely unknown. H...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi800067p
更新日期：2008-07-08 00:00:00
abstract：:Aminoglycoside 2''-phosphotransferase IVa [APH(2'')-IVa] is a member of a family of bacterial enzymes responsible for medically relevant resistance to antibiotics. APH(2'')-IVa confers high-level resistance against several clinically used aminoglycoside antibiotics in various pathogenic Enterococcus species by phospho...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi200747f
更新日期：2011-07-19 00:00:00
abstract：:The mitochondrial tyrosyl tRNA synthetase from Neurospora crassa (CYT-18 protein) is a bifunctional group I intron splicing cofactor. CYT-18 is capable of splicing multiple group I introns from a wide variety of sources by stabilizing the catalytically active intron structures. CYT-18 and mt TyrRSs from related fungal...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi401614h
更新日期：2014-03-04 00:00:00
abstract：:Trimethylamine dehydrogenase (TMADH) from the bacterium Methylophilus methylotrophus (sp. W(3)A(1)) and its C30A mutant were inactivated with three known inactivators of monoamine oxidase, namely, phenylhydrazine, N-cyclopropyl-alpha-methylbenzylamine, and 1-phenylcyclopropylamine. All three compounds irreversibly ina...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi0105126
更新日期：2001-07-24 00:00:00
abstract：:Transport of lactate, pyruvate, and the ketone bodies, acetoacetate and beta-hydroxybutyrate, is mediated in many mammalian cells by the monocarboxylate transporter MCT1. To be accepted as a substrate, a carboxyl group and an unpolar side chain are necessary. Site-directed mutagenesis of the rat MCT1 was used to ident...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi990973f
更新日期：1999-08-31 00:00:00
abstract：:β-Lactamase inhibitors (BLIs) restore the efficacy of otherwise obsolete β-lactams. However, commercially available BLIs are not effective against metallo-β-lactamases (MBLs), which continue to be disseminated globally. One group of the most clinically important MBLs is the VIM family. The discovery of VIM-24, a natur...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/acs.biochem.5b00106
更新日期：2015-05-26 00:00:00
abstract：:The reduction of substrates catalyzed by nitrogenase utilizes an electron transfer (ET) chain comprised of three metalloclusters distributed between the two component proteins, designated as the Fe protein and the MoFe protein. The flow of electrons through these three metalloclusters involves ET from the [4Fe-4S] clu...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi201003a
更新日期：2011-11-01 00:00:00
abstract：:The dependence of the activity of recombinant activated human protein C (r-APC) on each of its nine gamma-carboxyglutamic (Gla) residues (sequence positions 6, 7, 14, 16, 19, 20, 25, 26, and 29) has been assessed in purified systems and in plasma using r-mutants in which each Gla residue of r-APC was individually alte...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00173a033
更新日期：1994-02-22 00:00:00
abstract：:NADH oxidase activity has been detected in luminal membrane derived from the proximal tubules of rat kidney (brush border membranes). The cytochromes associated with these membranes [García, M.L., Benavides, J., Valdivieso, F., Mayor, F., & Giménez-Gallego, G. (1978) Biochem. Biophys. Res. Commun. 82, 738-744] were re...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00562a019
更新日期：1980-10-14 00:00:00
abstract：:Inheritance of apoE4 is a strong risk factor for the development of late-onset sporadic Alzheimer's disease (AD). Several lines of evidence suggest that apoE4 binds to the Alzheimer Abeta protein and, under certain experimental conditions, promotes formation of beta-sheet structures and amyloid fibrils. Deposition of ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi0114269
更新日期：2001-12-11 00:00:00
abstract：:Many current models of the Q cycle for the cytochrome (cyt) b6f and the cyt bc1 complexes incorporate 'Rieske' iron-sulfur protein (ISP) domain movements to gate electron transfer and to ensure high yields of proton shuttling. It was previously proposed that copper ions, which bind at a site distant from the quinol ox...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi015996k
更新日期：2002-03-26 00:00:00
abstract：:By using some mutant peptides of transcription factor Spl, phosphate backbone contacts with the DNA binding protein containing three zinc fingers have been investigated by alkylation interference, circular permutation, DNase I footprinting, and methylation protection methods. The ethylation interference analyses of Sp...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi952530r
更新日期：1996-07-02 00:00:00
abstract：:Casein kinase II (CKII) is a ubiquitous protein kinase, found predominantly in cell nuclei, which has two subunits in a tetrameric alpha 2 beta 2 or alpha alpha' beta 2 conformation. The catalytic center is present in the alpha subunit which is active by itself while beta is a regulatory subunit that can greatly enhan...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00079a030
更新日期：1993-07-20 00:00:00
