当前位置： SCI文献检索 > BIOCHEMISTRY期刊下所有文献 > Role of glycine 81 in (S)-mandelate dehydrogenase from Pseudomonas putida in substrate specificity and oxidase activity.

Role of glycine 81 in (S)-mandelate dehydrogenase from Pseudomonas putida in substrate specificity and oxidase activity.

Abstract：

:(S)-Mandelate dehydrogenase from Pseudomonas putida belongs to a FMN-dependent enzyme family that oxidizes (S)-alpha-hydroxyacids. Despite a high degree of sequence and structural similarity, this family can be divided into three subgroups based on the different oxidants utilized in the second oxidative half-reaction. Only the oxidases show high reactivity with molecular oxygen. Structural data indicate that the relative position of a peptide loop and the isoalloxazine ring of the FMN is slightly different in the oxidases compared to the dehydrogenases; the last residue on this loop is either an alanine or glycine. We examined the effect of the G81A, G81S, G81V, and G81D mutations in MDH on the overall reaction and especially on the suppression of activity with oxygen. G81A had a higher specificity for small substrates compared to that of wtMDH, though the affinity for (S)-mandelate was relatively unchanged. The rate of the first half-reaction was 20-130-fold slower for G81A and G81S; G81D and G81V had extremely low activity. Redox-potential measurements indicate that the reduction in activity is due to the decrease in electrophilicity of the FMN. The affinity for oxygen increased 10-15-fold for G81A and G81S relative to wtMDH; the rate of oxidation increased 2-fold for G81A. The increased reactivity with molecular oxygen did not correlate with the redox potentials and appears to primarily result from a higher affinity for oxygen. These results suggest that one of the ways the oxidase activity of MDH is controlled is through steric effects because of the relative positions of the FMN and the Gly81 loop.

journal_name

Biochemistry

journal_title

Biochemistry

authors

Dewanti AR,Xu Y,Mitra B

doi

10.1021/bi049005p

subject

Has Abstract

pub_date

2004-08-24 00:00:00

pages

10692-700

issue

33

eissn

0006-2960

issn

1520-4995

journal_volume

43

pub_type

杂志文章

相关文献

BIOCHEMISTRY文献大全
  • Substrate binding to the peripheral site of acetylcholinesterase initiates enzymatic catalysis. Substrate inhibition arises as a secondary effect.

    abstract：:Two sites of ligand interaction in acetylcholinesterase (AChE) were first demonstrated in ligand binding studies and later confirmed by crystallography, site-specific mutagenesis, and molecular modeling: an acylation site at the base of the active site gorge and a peripheral site at its mouth. We recently introduced a...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi9813577

    authors： Szegletes T,Mallender WD,Thomas PJ,Rosenberry TL

    更新日期：1999-01-05 00:00:00

  • Nanosecond fluorescence studies of noncovalent interaction of monomeric and dimeric intercalators with DNA.

    abstract：:The noncovalent interaction of 2-aminodipyrido[1,2-a:3',2'-d]imidazole (Glu-P-2) and its derivatives, which are potent mutagens isolated from L-glutamic acid pyrolysate, with calf thymus DNA was studied by steady-state and nanosecond fluorescence spectroscopies. The fluorescence of these compounds exhibits static quen...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00344a013

    authors： Itoh M,Kurokawa H,Usui M,Ohno M,Shimoda E,Hashimoto Y,Shudo K

    更新日期：1985-11-05 00:00:00

  • Structural and functional analysis of the homing endonuclease PI-sceI by limited proteolytic cleavage and molecular cloning of partial digestion products.

    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

    authors： Pingoud V,Grindl W,Wende W,Thole H,Pingoud A

    更新日期：1998-06-02 00:00:00

  • Cloning and expression of three rabbit kidney cDNAs encoding lauric acid omega-hydroxylases.

    abstract：:cDNAs encoding three cytochrome P-450 enzymes were cloned from a rabbit kidney cDNA library. These three cDNAs exhibit greater than 90% nucleotide sequence identity across the coding region. This degree of sequence identity is also seen with P450IVA4, an enzyme that catalyzes the omega-hydroxylation of prostaglandins ...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00456a004

    authors： Johnson EF,Walker DL,Griffin KJ,Clark JE,Okita RT,Muerhoff AS,Masters BS

    更新日期：1990-01-30 00:00:00

  • The reactivity of heme in biological systems: autocatalytic formation of both tyrosine-heme and tryptophan-heme covalent links in a single protein architecture.

    abstract：:We have previously shown that introduction of an engineered Met160 residue in ascorbate peroxidase (S160M variant) leads to the formation of a covalent link between Met160 and the heme vinyl group [Metcalfe, C. L., et al. (2004) J. Am. Chem. Soc. 126, 16242-16248]. In this work, we have used electronic spectroscopy, H...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi7015316

    authors： Pipirou Z,Bottrill AR,Svistunenko DA,Efimov I,Basran J,Mistry SC,Cooper CE,Raven EL

    更新日期：2007-11-20 00:00:00

  • A transition state analogue for two pyruvate metabolizing enzymes, lactate dehydrogenase and alanine dehydrogenase.

    abstract：:The synthesis of 5-(2-oxalylethyl)-NADH, a reduced nicotinamide adenine dinucleotide (NADH) derivate with pyruvate covalently attached to the 5 position of the dihydronicotinamide ring over an additional methylene group has been described previously (Trommer, W.E., Blume, H., and Kapmeyer, H. (1976) Justus Liebigs Ann...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00668a012

    authors： Kapmeyer H,Pfleiderer G,Trommer WE

    更新日期：1976-11-16 00:00:00

  • A method for assessing the stability of a membrane protein.

    abstract：:The integral membrane protein diacylglycerol kinase (DGK) from Escherichia coli has been reversibly unfolded in a protein/detergent/mixed micelle system by varying the molar ratio of n-decyl beta-D-maltoside (DM) and sodium dodecyl sulfate (SDS). Unfolding was monitored by circular dichroism (CD) and ultraviolet (UV) ...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi963095j

    authors： Lau FW,Bowie JU

    更新日期：1997-05-13 00:00:00

  • Distal site aspartate is essential in the catalase activity of catalase-peroxidases.

    abstract：:Structural and biochemical characterization of aspartate 152 at the distal heme side of catalase-peroxidase (KatG) from Synechocystis PCC 6803 reveals an important functional role for this residue. In the wild-type protein, the side chain carboxyl group of Asp152 is 7.8 A apart from the heme iron and is hydrogen-bonde...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi026944d

    authors： Jakopitsch C,Auer M,Regelsberger G,Jantschko W,Furtmüller PG,Rüker F,Obinger C

    更新日期：2003-05-13 00:00:00

  • Toward an understanding of the mechanism of nonphotochemical quenching in green plants.

    abstract：:Oxygenic photosynthesis in plants involves highly reactive intermediates and byproducts that can damage the photosynthetic apparatus and other chloroplast constituents. The potential for damage is exacerbated when the amount of absorbed light exceeds the capacity for light energy utilization in photosynthesis, a condi...

    journal_title：Biochemistry

    pub_type： 杂志文章,评审

    doi：10.1021/bi0494020

    authors： Holt NE,Fleming GR,Niyogi KK

    更新日期：2004-07-06 00:00:00

  • DNA sequence-specific recognition by the Saccharomyces cerevisiae "TATA" binding protein: promoter-dependent differences in the thermodynamics and kinetics of binding.

    abstract：:The equilibrium binding and association kinetics of the Saccharomyces cerevisiae TATA Binding Protein (TBP) to the E4 and Major Late promoters of adenovirus (TATATATA and TATAAAAG, respectively), have been directly compared by quantitative DNase I titration and quench-flow "footprinting". The equilibrium binding of TB...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi981072u

    authors： Petri V,Hsieh M,Jamison E,Brenowitz M

    更新日期：1998-11-10 00:00:00

  • Solution structure of decorin-binding protein A from Borrelia burgdorferi.

    abstract：:Decorin-binding protein A (DBPA) is an important lipoprotein from the bacterium Borrelia burgdorferi, the causative agent of Lyme disease. The absence of DBPA drastically reduces the pathogenic potential of the bacterium, and biochemical evidence indicates DBPA's interactions with the glycosaminoglycan (GAG) portion o...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi3007093

    authors： Wang X

    更新日期：2012-10-23 00:00:00

  • Role of GTP hydrolysis in microtubule polymerization: evidence for a coupled hydrolysis mechanism.

    abstract：:The relationship between GTP hydrolysis and microtubule assembly has been investigated by using a rapid filtration method. Microtubules assembled from phosphocellulose-purified tubulin, double-labeled with [gamma-32P]- and [3H]GTP, were trapped and washed free of unbound nucleotide on glass fiber filters. The transien...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00479a022

    authors： Stewart RJ,Farrell KW,Wilson L

    更新日期：1990-07-10 00:00:00

  • Duplicated dockerin subdomains of Clostridium thermocellum endoglucanase CelD bind to a cohesin domain of the scaffolding protein CipA with distinct thermodynamic parameters and a negative cooperativity.

    abstract：:Mutagenized dockerin domains of endoglucanase CelD (type I) and of the cellulosome-integrating protein CipA (type II) were constructed by swapping residues 10 and 11 of the first or the second duplicated segment between the two polypeptides. These residues have been proposed to determine the specificity of cohesin-doc...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi011853m

    authors： Schaeffer F,Matuschek M,Guglielmi G,Miras I,Alzari PM,Béguin P

    更新日期：2002-02-19 00:00:00

  • A unified kinetic mechanism applicable to multiple DNA polymerases.

    abstract：:After extensive studies spanning over half a century, there is little consensus on the kinetic mechanism of DNA polymerases. Using stopped-flow fluorescence assays for mammalian DNA polymerase beta (Pol beta), we have previously identified a fast fluorescence transition corresponding to conformational closing, and a s...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi700084w

    authors： Bakhtina M,Roettger MP,Kumar S,Tsai MD

    更新日期：2007-05-08 00:00:00

  • NH2-terminal extensions on skin collagen from sheep with a genetic defect in conversion of procollagen into collagen.

    abstract：:A modified form of procollagen was extracted with 10 M urea from the skin of lambs with dermatosparaxis, a disease which is produced by a genetic defect in the conversion of procollagen to collagen. The extracts contained little if any alpha1 and alpha2 chains of normal type I collagen, and instead they contained the ...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00658a024

    authors： Becker U,Timpl R

    更新日期：1976-06-29 00:00:00

  • Complex of human apolipoprotein C-1 with phospholipid: thermodynamic or kinetic stability?

    abstract：:Thermal unfolding of discoidal complexes of apolipoprotein (apo) C-1 with dimyristoyl phosphatidylcholine (DMPC) reveals a novel mechanism of lipoprotein stabilization that is based on kinetics rather than thermodynamics. Far-UV CD melting curves recorded at several heating/cooling rates from 0.047 to 1.34 K/min show ...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi025588w

    authors： Gursky O,Ranjana,Gantz DL

    更新日期：2002-06-11 00:00:00

  • Dissociation of the α-subunit Calf-2 domain and the β-subunit I-EGF4 domain in integrin activation and signaling.

    abstract：:Integrin conformational changes mediate integrin activation and signaling triggered by intracellular molecules or extracellular ligands. Even though it is known that αβ transmembrane domain separation is required for integrin signaling, it is still not clear how this signal is transmitted from the transmembrane domain...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi101462h

    authors： Wang W,Fu G,Luo BH

    更新日期：2010-11-30 00:00:00

  • Uracil DNA-glycosylase/glyceraldehyde-3-phosphate dehydrogenase is an Ap4A binding protein.

    abstract：:A 37 kDa protein that binds to diadenosine tetraphosphate (Ap4A) was purified from human HeLa cells and identified as uracil DNA glycosylase/glyceraldehyde-3-phosphate dehydrogenase (UDG/GAPDH). Utilizing photoaffinity labeling with [alpha-32P]8N3-Ap4A, an Ap4A binding protein of 37 kDa was identified from HeLa cell n...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00030a007

    authors： Baxi MD,Vishwanatha JK

    更新日期：1995-08-01 00:00:00

  • Gene heterogeneity: a basis for alternative 5.8S rRNA processing.

    abstract：:Two bands of 5.8S rRNA were observed when the total RNA isolated from rat or mouse tissue was separated by electrophoresis on high-resolution polyacrylamide gels under denaturing conditions. The minor form, with a lower mobility, represented 15-35% of the total 5.8S rRNA, depending on the source of the tissue. Sequenc...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00311a011

    authors： Smith SD,Banerjee N,Sitz TO

    更新日期：1984-07-31 00:00:00

  • Operative Binding of Class I Release Factors and YaeJ Stabilizes the Ribosome in the Nonrotated State.

    abstract：:During translation, the small subunit of the ribosome rotates with respect to the large subunit primarily between two states as mRNA is being translated into a protein. At the termination of bacterial translation, class I release factors (RFs) bind to a stop codon in the A-site and catalyze the release of the peptide ...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/acs.biochem.7b00824

    authors： Casy W,Prater AR,Cornish PV

    更新日期：2018-04-03 00:00:00

  • N-ethylmaleimide inhibits Ncd motor function by modification of a cysteine in the stalk domain.

    abstract：:N-Ethylmaleimide (NEM), which reacts readily with exposed sulfhydryl groups, has been shown to inhibit the activity of the microtubule (MT) motors kinesin, Ncd, and dynein. Currently, the mechanism of inhibition is not known for any of these proteins. To investigate the mechanism by which NEM inhibits Ncd, the recombi...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi990706+

    authors： Phelps KK,Walker RA

    更新日期：1999-08-17 00:00:00

  • Structural behavior of the CpG step in two related oligonucleotides reflects its malleability in solution.

    abstract：:We report on the determination of the solution structure of two sequence-related oligonucleotides, d(GTACGTAC)2 and d(CATCGATG)2. Results have been obtained by using a combined approach of (a) two-dimensional NMR, including proton and phosphorus experiments, (b) restrained molecular mechanics performed with sugar phas...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00037a045

    authors： Lefebvre A,Mauffret O,Hartmann B,Lescot E,Fermandjian S

    更新日期：1995-09-19 00:00:00

  • Oxidative stress induced by a copper-thiosemicarbazone complex.

    abstract：:Copper thiosemicarbazones cause considerable oxidative stress. This effect may be related to their cytotoxicity. In the present work, the chemical and cellular properties of a new ligand, pyridoxal thiosemicarbazone (H2T), and its copper(II) chelate (CuT) are assessed. CuT is toxic to cultured Ehrlich ascites tumor ce...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00482a014

    authors： Byrnes RW,Mohan M,Antholine WE,Xu RX,Petering DH

    更新日期：1990-07-31 00:00:00

  • Study of hydrogen exchange in hemoglobin as a function of fractional ligand saturation.

    abstract：:We have studied the hydrogen-exchange kinetics of hemoglobin A0 as a function of ligand, CO, saturation. In the noncooperative system, azide binding to methemoglobin, the alterations in exchange kinetics are proportional to the average degree of ligation. However, in the case of CO binding to deoxyhemoglobin the chang...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00297a015

    authors： Hallaway PE,Hallaway BE,Rosenberg A

    更新日期：1984-01-17 00:00:00

  • Alternate substrate binding modes to two mutant (D98N and H255N) forms of nitrite reductase from Alcaligenes faecalis S-6: structural model of a transient catalytic intermediate.

    abstract：:High-resolution nitrite soaked oxidized and reduced crystal structures of two active site mutants, D98N and H255N, of nitrite reductase (NIR) from Alcaligenes faecalis S-6 were determined to better than 2.0 A resolution. In the oxidized D98N nitrite-soaked structures, nitrite is coordinated to the type II copper via i...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi0107400

    authors： Boulanger MJ,Murphy ME

    更新日期：2001-08-07 00:00:00

  • Addition of lysines to the 50/20 kDa junction of myosin strengthens weak binding to actin without affecting the maximum ATPase activity.

    abstract：:Much interest has centered on two surface loops in the motor domain to explain the differences in enzymatic and mechanical properties of myosin isoforms. We showed that two invariant lysines at the C-terminal end of loop 2, which is part of the actin-binding interface, are required to obtain actin activation [Joel et ...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi034415j

    authors： Joel PB,Sweeney HL,Trybus KM

    更新日期：2003-08-05 00:00:00

  • Thermodynamics of nucleotide and inhibitor binding to wild-type and ispinesib-resistant forms of human kinesin spindle protein.

    abstract：:Current antimitotic cancer chemotherapy based on vinca alkaloids and taxanes target tubulin, a protein required not only for mitotic spindle formation but also for the overall structural integrity of terminally differentiated cells. Among many innovations targeting specific mitotic events, inhibition of motor enzymes ...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi900946r

    authors： Sheth PR,Basso A,Duca JS,Lesburg CA,Ogas P,Gray K,Nale L,Mannarino AF,Prongay AJ,Le HV

    更新日期：2009-11-24 00:00:00

  • Differential regulation of WASP and N-WASP by Cdc42, Rac1, Nck, and PI(4,5)P2.

    abstract：:The Wiskott-Aldrich syndrome protein (WASP) and neural WASP (N-WASP) are key players in regulating actin cytoskeleton via the Arp2/3 complex. It has been widely reported that the WASP proteins are activated by Rho family small GTPase Cdc42 and that Rac1 acts through SCAR/WAVE proteins. However, a systematic study of t...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi062152y

    authors： Tomasevic N,Jia Z,Russell A,Fujii T,Hartman JJ,Clancy S,Wang M,Beraud C,Wood KW,Sakowicz R

    更新日期：2007-03-20 00:00:00

  • Thermodynamic characterization of naturally occurring RNA single mismatches with G-U nearest neighbors.

    abstract：:Due to their prevalence and roles in biological systems, single mismatches adjacent to G-U pairs are important RNA structural elements. Since there are only limited experimental values for the stability of single mismatches adjacent to G-U pairs, current algorithms using free energy minimization to predict RNA seconda...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi800471z

    authors： Davis AR,Znosko BM

    更新日期：2008-09-23 00:00:00

  • Magnetic circular dichroism properties of reaction center complexes isolated from the zinc-bacteriochlorophyll a-containing purple bacterium Acidiphilium rubrum.

    abstract：:Reaction center (RC) complexes isolated from a Zn-bacteriochlorophyll (BChl) a-containing purple bacterium, Acidiphilium rubrum, were characterized by absorption, circular dichroism, and magnetic circular dichroism (MCD) spectroscopy. The oxidized-minus-reduced difference spectra indicated that, in this RC, the Zn-BCh...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi991444e

    authors： Mimuro M,Kobayashi M,Shimada K,Uezono K,Nozawa T

    更新日期：2000-04-11 00:00:00