Abstract：
:To determine the active site cysteine residue in aldehyde dehydrogenase, we mutated amino acid residues 49, 162, and 302 of recombinantly expressed rat liver mitochondrial (class 2) aldehyde dehydrogenase. The C49A and C162A mutants were fully active tetrameric enzymes, although the C162A mutant was found to be highly unstable. The C302A mutant was also a tetramer and bound coenzyme, but lacked both dehydrogenase and esterase activities. To test for the role of cysteine 302 as a nucleophile, the residue was mutated to a serine, a poor nucleophile. this C302S mutant was active but was a much poorer catalyst, with a kcat/Km value 7 x 10(5) times lower than that of the recombinant native enzyme. Unlike with native enzyme where deacylation is rate limiting, formation of the serine hemiacetal intermediate appeared to be the rate-limiting step. Cysteine 302 is the only strictly conserved cysteine residue among all the available sequences of the aldehyde dehydrogenase superfamily, supporting the role of this residue as the active site nucleophile of aldehyde dehydrogenase.
journal_name
Biochemistryjournal_title
Biochemistryauthors
Farrés J,Wang TT,Cunningham SJ,Weiner Hdoi
10.1021/bi00008a025subject
Has Abstractpub_date
1995-02-28 00:00:00pages
2592-8issue
8eissn
0006-2960issn
1520-4995journal_volume
34pub_type
杂志文章相关文献
BIOCHEMISTRY文献大全abstract：:We have previously suggested that folding of the SV40 attenuator RNA 1 into two hairpin elements leads to a block of transcription elongation. Using site-directed mutagenesis, we created three templates that either strengthened or weakened the proposed hairpin structures. The mutated and wild-type templates were clone...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00150a034
更新日期：1992-09-08 00:00:00
abstract：:Quinone oxidoreductase 2 (QR2) purified from human red blood cells was recently shown to be a potential target of the quinoline antimalarial compounds [Graves et al., (2002) Mol. Pharmacol. 62, 1364]. QR2 catalyzes the two-electron reduction of menadione via the oxidation of N-alkylated or N-ribosylated nicotinamides....
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi035923w
更新日期：2004-04-20 00:00:00
abstract：:Polyisoprenyl diphosphates play diverse and vital roles in cell function in health and disease. The counter-regulatory lipid signaling molecule, presqualene diphosphate (PSDP), is rapidly converted to its monophosphate form (PSMP) upon cell activation [Levy, B. D., Petasis, N. A., and Serhan, C. N. (1997) Nature 389, ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi8020636
更新日期：2009-04-07 00:00:00
abstract：:This study presents an approach to identifying surface residues on membrane proteins that are exposed toward the membrane-aqueous interface. The method employs a lipid Ni(II) chelate that localizes the metal ion to a region near the membrane-aqueous interface. Lateral diffusion of the lipid chelate results in Heisenbe...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi015828s
更新日期：2002-01-29 00:00:00
abstract：:In two recent papers, we reported the effects of several point mutations on the thermodynamics of the thermal unfolding of the lysozyme of phage T4 as determined by differential scanning calorimetry. The mutants studied were R96H [Kitamura, S., & Sturtevant, J.M. (1989) Biochemistry 28, 3788-3792] and T157 replaced by...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00121a009
更新日期：1992-02-18 00:00:00
abstract：:The heat-stable, protein inhibitor of the cyclic adenosine monophosphate (cAMP) dependent protein kinase [Walsh, D. A., Ashby, C. D., Gonzalez, C., Calkins, D., Fischer, E., & Krebs, E (1971a) J. Biol. Chem. 246, 1977-1985] has been purified to homogeneity from rabbit skeletal muscle by preparative electrophoresis. Em...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00589a011
更新日期：1979-10-30 00:00:00
abstract：:Dynamic conformations of two distinct immunoglobulin (Ig) isotypes, murine IgE and human IgG1, were examined with fluorescence resonance energy transfer measurements. The IgE mutant epsilon/C gamma 3* and the IgG1 mutant gamma/C gamma 3* each bind [5-(dimethylamino)naphthalen-1-yl]sulfonyl (DNS) in two identical antig...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00148a004
更新日期：1992-08-25 00:00:00
abstract：:The role of lysines in the ribonucleolytic and angiogenic activities of human angiogenin has been examined by chemical modification and site-directed mutagenesis. It was demonstrated previously [Shapiro, R., Weremowicz, S., Riordan, J.F., & Vallee, B.L. (1987) Proc. Natl. Acad. Sci. U.S.A. 84, 8783-8787] that extensiv...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00430a045
更新日期：1989-02-21 00:00:00
abstract：:The genome of all retroviruses consists of two identical RNAs noncovalently linked near their 5' end. Dimerization of genomic RNA is thought to modulate several steps in the retroviral life cycle, such as recombination, translation, and encapsidation. The kissing-loop model of HIV-1 genome dimerization [Laughrea, M., ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi951838f
更新日期：1996-02-06 00:00:00
abstract：:The membrane-bound quinoprotein glucose dehydrogenase (mGDH) in Escherichia coli contains pyrroloquinoline quinone (PQQ) and participates in the direct oxidation of D-glucose to D-gluconate by transferring electrons to ubiquinone (UQ). To elucidate the mechanism of ubiquinone reduction by mGDH, we applied a pulse radi...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi051347n
更新日期：2005-10-18 00:00:00
abstract：:The properties of Na,K-ATPase phosphoenzymes formed either from ATP in the presence of Mg2+ and Na+ or from Pi in the absence of alkali cations were investigated by biochemical methods and spectrofluorometry employing the styryl dye RH421. We characterized the phosphoenzyme species by their reaction to N-methyl hydrox...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi980703h
更新日期：1998-09-29 00:00:00
abstract：:We have investigated in detail the cooperativity which we had previously observed in the pH titration profiles of the histidine residues of rabbit tropomyosin [Edwards, B. F. P., & Sykes, B. D. (1978) Biochemistry 17, 684]. Nonpolymerizing tropomyosin was prepared by carboxypeptidase digestion, and the titration profi...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00517a037
更新日期：1981-07-07 00:00:00
abstract：:The compounds 3-hydroxy-4-nitrobenzaldehyde and 3-hydroxy-4-nitrobenzyl alcohol are introduced as new chromophoric substrates for probing the catalytic mechanism of horse liver alcohol dehydrogenase (LADH). Ionization of the phenolic hydroxyl group shifts the spectrum of the aldehyde from 360 to 433 nm (pKa = 6.0), wh...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00385a017
更新日期：1987-06-02 00:00:00
abstract：:Conjugative transfer of F-like plasmids in Escherichia coli is repressed by a plasmid-encoded protein, FinO. FinO blocks the translation of TraJ, a positive activator of transcription of genes required for conjugation. FinO binds a traJ antisense RNA, FinP, thereby protecting it from degradation, and catalyzes FinP-tr...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi9911482
更新日期：1999-10-19 00:00:00
abstract：:Tripeptidyl peptidase II is a high molecular weight serine exopeptidase, which has been purified from rat liver and human erythrocytes. Four clones, representing 4453 bp, or 90% of the mRNA of the human enzyme, have been isolated from two different cDNA libraries. One clone, designated A2, was obtained after screening...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00215a025
更新日期：1991-01-08 00:00:00
abstract：:We have characterized by EPR the interaction of the Em,7 = -50 mV [4Fe-4S] cluster of Escherichia coli DMSO reductase (DmsABC) with a menaquinol (MQH2) binding site. Potentiometric titrations indicate that in DmsAB(C102S)C, the Em,7 = -50 mV [4Fe-4S] cluster is replaced by an Em,7 = +260 mV [3Fe-4S] cluster. The Q-poo...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi951584y
更新日期：1996-03-12 00:00:00
abstract：:Extracellular cAMP induces the rapid activation of guanylate cyclase, which adapts within 10 s to constant cAMP concentrations. A new response can be induced either by a higher cAMP concentration or by the same cAMP concentration at some time (t1/2 = 90 s) after removal of the previous stimulus. Stimulation of guanyla...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00397a047
更新日期：1987-11-17 00:00:00
abstract：:All-trans-retinol is the precursor for all-trans-retinoic acid, the activating ligand for nuclear transcription factors retinoic acid receptors. In the cytosol of various cells, most retinol exists in a bound form, complexed with cellular retinol binding protein type I (holo-CRBP). Whether retinoic acid is produced fr...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi026836r
更新日期：2003-01-28 00:00:00
abstract：:Self-splicing of Tetrahymena pre-rRNA proceeds in two consecutive phosphoryl transesterification steps. One major difference between these steps is that in the first an exogenous guanosine (G) binds to the active site, while in the second the 3'-terminal G414 residue of the intron binds. The first step has been extens...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi9527653
更新日期：1996-05-07 00:00:00
abstract：:A method is described for using very high specific activity [3H]poly(deoxythymidylate) [[3H]poly(dT)] to detect, size, and quantiate subnanogram amounts of nonradioactive polyadenylated RNA. Short (approximately 100 nucleotides long) [3H]poly(dT) is hybridized to the poly(adenylate) [poly(A)] tracts in polyadenylated ...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00557a022
更新日期：1980-08-05 00:00:00
abstract：:PDZ (acronym of the synapse-associated protein PSD-95/SAP90, the septate junction protein Discs-large, and the tight junction protein ZO-1) domains are abundant small globular protein interaction domains that mainly recognize the carboxyl termini of their target proteins. Detailed knowledge on PDZ domain binding speci...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi700954e
更新日期：2007-11-27 00:00:00
abstract：:The baker's yeast saccharopine dehydrogenase (EC 1.5.1.7) was inactivated by 2,3-butanedione following pseudo-first-order reaction kinetics. The pseudo-first-order rate constant for inactivation was linearly related to the butanedione concentration, and a value of 7.5 M-1 min-1 was obtained for the second-order rate c...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00506a004
更新日期：1981-02-03 00:00:00
abstract：:The photocycle of 124 kDa phytochrome A from Avena sativa was studied by Fourier-transform infrared spectroscopy at low temperatures. Difference spectra between the parent state Pr and the intermediates of the Pr-->Pfr pathway, i.e. lumi-R, meta-Ra, and meta-Rc, and between Pfr and the intermediates of the Pfr-->Pr pa...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi960960r
更新日期：1996-08-20 00:00:00
abstract：:Under suitable experimental conditions the aerobic oxidation of NADH catalyzed by horseradish peroxidase occurred in four characteristic phases: initial burst, induction phase, steady state, and termination. A trace amount of H2O2 present in the NADH solution brought about initial burst in the formation of oxyperoxida...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00628a024
更新日期：1977-05-03 00:00:00
abstract：:A single G3:U70 base pair in the acceptor helix is the major determinant of alanine acceptance in alanine transfer RNAs. Transfer of this base pair into other transfer RNAs confers alanine acceptance. A G3:C70 substitution eliminates alanine acceptance in vivo and in vitro. In this work, a population of mutagenized G3...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi00157a019
更新日期：1992-10-27 00:00:00
abstract：:myo-Inositol oxygenase (MIOX) uses iron as its cofactor and dioxygen as its cosubstrate to effect the unique, ring-cleaving, four-electron oxidation of its cyclohexan-(1,2,3,4,5,6-hexa)-ol substrate to d-glucuronate. The nature of the iron cofactor and its interaction with the substrate, myo-inositol (MI), have been p...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi0519607
更新日期：2006-05-02 00:00:00
abstract：:In Bacillus subtilis, P protein is the noncatalytic component of ribonuclease P (RNase P) that is critical for achieving maximal nuclease activity under physiological conditions. P protein is predominantly unfolded (D) at neutral pH and low ionic strength; however, it folds upon the addition of sulfate anions (ligands...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi0504613
更新日期：2005-10-04 00:00:00
abstract：:The aldehyde oxidoreductase from Desulfovibrio gigas belongs to the family of molybdenum hydroxylases. Besides a molybdenum cofactor which constitutes their active site, these enzymes contain two [2Fe-2S](2+,1+) clusters which are believed to transfer the electrons provided by the substrate to an acceptor which is eit...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi0510025
更新日期：2005-08-30 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：:Multiply branched polylactosaminoglycans are expressed in glycoproteins and glycolipids of many cells. Interest in their biology stems from their abundant expression in early embryonal cells and from their ability to carry multiple lectin-binding determinants, which makes them prominent ligands and antagonists of cell...
journal_title：Biochemistry
pub_type： 杂志文章
doi：10.1021/bi9627673
更新日期：1997-06-10 00:00:00
