当前位置： SCI文献检索 > BIOCHEMISTRY期刊下所有文献 > Active site modifications in a double mutant of liver alcohol dehydrogenase: structural studies of two enzyme-ligand complexes.

Active site modifications in a double mutant of liver alcohol dehydrogenase: structural studies of two enzyme-ligand complexes.

Abstract：

:The oxidation of alcohol to aldehyde by horse liver alcohol dehydrogenase (LADH) requires the transfer of a hydride ion from the alcohol substrate to the cofactor nicotinamide adenine dinucleotide (NAD). A quantum mechanical tunneling contribution to this hydride transfer step has been demonstrated in a number of LADH mutants designed to enhance or diminish this effect [Bahnson, B. J., et al. (1997) Proc. Natl. Acad. Sci. U.S.A. 94, 12797-12802]. The active site double mutant Phe93 --> Trp/Val203 --> Ala shows a 75-fold reduction in catalytic efficiency relative to that of the native enzyme, and reduced tunneling relative to that of either single mutant. We present here two crystal structures of the double mutant: a 2.0 A complex with NAD and the substrate analogue trifluoroethanol and a 2.6 A complex with the isosteric NAD analogue CPAD and ethanol. Changes at the active site observed in both complexes are consistent with reduced activity and tunneling. The NAD-trifluoroethanol complex crystallizes in the closed conformation characteristic of the active enzyme. However, the NAD nicotinamide ring rotates away from the substrate, toward the space vacated by replacement of Val203 with the smaller alanine. Replacement of Phe93 with the larger tryptophan also produces unfavorable steric contacts with the nicotinamide carboxamide group, potentially destabilizing hydrogen bonds required to maintain the closed conformation. These contacts are relieved in the second complex by rotation of the CPAD pyridine ring into an unusual syn orientation. The resulting loss of the carboxamide hydrogen bonds produces an open conformation characteristic of the apoenzyme.

journal_name

Biochemistry

journal_title

Biochemistry

authors

Colby TD,Bahnson BJ,Chin JK,Klinman JP,Goldstein BM

doi

10.1021/bi973184b

subject

Has Abstract

pub_date

1998-06-30 00:00:00

pages

9295-304

issue

26

eissn

0006-2960

issn

1520-4995

pii

bi973184b

journal_volume

37

pub_type

杂志文章

相关文献

BIOCHEMISTRY文献大全
  • DNA Oligonucleotide 3'-Phosphorylation by a DNA Enzyme.

    abstract：:T4 polynucleotide kinase is widely used for 5'-phosphorylation of DNA and RNA oligonucleotide termini, but no natural protein enzyme is capable of 3'-phosphorylation. Here, we report the in vitro selection of deoxyribozymes (DNA enzymes) capable of DNA oligonucleotide 3'-phosphorylation, using a 5'-triphosphorylated R...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/acs.biochem.6b00151

    authors： Camden AJ,Walsh SM,Suk SH,Silverman SK

    更新日期：2016-05-10 00:00:00

  • 6-(Methoxymethylene)penicillanic acid: inactivator of RTEM beta-lactamase from Escherichia coli.

    abstract：:The Z and E isomers of 6-(methoxymethylene)-penicillanic acid have been synthesized, and their interaction with the RTEM beta-lactamase has been studied. The Z isomer is an inhibitor and an inactivator of the enzyme, and there is some similarity between its behavior and that of other mechanism-based inactivators such ...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00319a025

    authors： Brenner DG,Knowles JR

    更新日期：1984-11-20 00:00:00

  • Influence of the extent of branching on solution conformations of complex oligosaccharides: a molecular dynamics and NMR study of a penta-antennary "bisected" N-glycan.

    abstract：:The solution conformation of an agalactosyl penta-antennary "bisected" N-linked glycan from hen ovomucoid has been determined using a combination of 1H-NMR NOE measurements and restrained molecular dynamics (MD) simulations. The majority of glycosidic linkages exhibited restricted torsional fluctuations about the glob...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00043a018

    authors： Rutherford TJ,Neville DC,Homans SW

    更新日期：1995-10-31 00:00:00

  • Isolation and characterization of an organic solvent soluble polypeptide component from photoreceptor complexes of Rhodospirillum rubrum.

    abstract：:An organic solvent soluble polypeptide has been isolated from photoreceptor complexes and chromatophores of Rhodospirillum rubrum. After extraction of the protein from lyophilized samples with 1:1 chloroform-methanol, it was purified by column chromatography. Its isoelectric point determined by isoelectric focusing wa...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00624a011

    authors： Tonn SJ,Gogel GE,Loach PA

    更新日期：1977-03-08 00:00:00

  • Major groove (R)-alpha-(N6-adenyl)styrene oxide adducts in an oligodeoxynucleotide containing the human N-ras codon 61 sequence: conformations of the R(61,2) and R(61,3) sequence isomers from 1H NMR.

    abstract：:Conformations of (R)-alpha-(N6-adenyl)styrene oxide adducts at positions X6 in d(CGGACXAGAAG).d(CTTCTTGTCCG) and X7 in d(CGGACAXGAAG).d(CTTCTTGTCCG), incorporating codons 60, 61 (underlined), and 62 of the human n-ras protooncogene, were refined from 1H NMR data. These were the R(61,2) and R(61,3) adducts. Chemical sh...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00043a008

    authors： Feng B,Zhou L,Passarelli M,Harris CM,Harris TM,Stone MP

    更新日期：1995-10-31 00:00:00

  • Hydrolysis of plant cuticle by plant pathogens. Properties of cutinase I, cutinase II, and a nonspecific esterase isolated from Fusarium solani pisi.

    abstract：:The properties of the homogeneous cutinase I, cutinase II, and the nonspecific esterase isolated from the extracellular fluid of cutin-grown Fusarium solani F. pisi (R.E. Purdy and P.E. Kolattukudy (1975), Biochemistry, preceding paper in this issue) were investigated. Using tritiated apple cutin as substrate, the two...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00684a007

    authors： Purdy RE,Kolattukudy PE

    更新日期：1975-07-01 00:00:00

  • Light-chain-independent binding of adaptors, AP180, and auxilin to clathrin.

    abstract：:Binding of coated vesicle assembly proteins to clathrin causes it to assemble into regular coat structures. The assembly protein fraction of bovine brain coated vesicles comprises AP180, auxilin, and HA1 and HA2 adaptors. Clathrin heavy chains, separated from their light chains, polymerize with unimpaired efficiency w...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00101a027

    authors： Lindner R,Ungewickell E

    更新日期：1991-09-17 00:00:00

  • Primary structure of human J chain: alignment of peptides from chemical and enzymatic hydrolyses.

    abstract：:The primary structure of the J chain from a human Waldenströms IgM protein has been determined using a combination of automated and conventional Edman degradative procedures. Eighty-five percent of the sequence was established with peptides isolated from tryptic digests of carboxyamidomethylated and citraconylated J c...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00635a002

    authors： Mole JE,Bhown AS,Bennett JC

    更新日期：1977-08-09 00:00:00

  • Copper binding to beta-2-microglobulin and its pre-amyloid oligomers.

    abstract：:Beta-2-microglobulin (beta2m) deposits as amyloid fibrils in the musculoskeletal system of patients undergoing long-term dialysis treatment as a result of kidney failure. Previous work has shown that Cu(II) binding causes beta2m to organize into nativelike dimers and tetramers that precede amyloid formation. Cu(II) is...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi901172y

    authors： Srikanth R,Mendoza VL,Bridgewater JD,Zhang G,Vachet RW

    更新日期：2009-10-20 00:00:00

  • Characterization of the ATPase activity of purified Chinese hamster P-glycoprotein.

    abstract：:A simple and rapid procedure is described for purification of P-glycoprotein (Pgp) from a multidrug-resistant Chinese hamster ovary cell line (CR1R12) in which the plasma membranes are highly enriched in Pgp (Al-Shawi, M.K., Senior A.E. (1993) J. Biol. Chem, 268, 4197-4206). The procedure consisted of octylglucoside s...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00189a008

    authors： Urbatsch IL,al-Shawi MK,Senior AE

    更新日期：1994-06-14 00:00:00

  • Heterologous expression of photoreceptor peripherin/rds and Rom-1 in COS-1 cells: assembly, interactions, and localization of multisubunit complexes.

    abstract：:Peripherin/rds is a 39 kDa integral membrane glycoprotein essential for normal photoreceptor cell development in vertebrates. It has been implicated in several human retinal degenerative diseases including retinitis pigmentosa and macular degeneration and is thought to play a structural role at photoreceptor outer seg...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00043a028

    authors： Goldberg AF,Moritz OL,Molday RS

    更新日期：1995-10-31 00:00:00

  • Preferential assembly of the tropomyosin heterodimer: equilibrium studies.

    abstract：:Thermal unfolding/refolding studies of the three tropomyosin dimers, alpha alpha, alpha beta, and beta beta, from chicken gizzard muscle were performed to explain the preferential assembly of alpha- and beta-tropomyosin subunits into heterodimers, alpha beta [Lehrer, S. S., & Qian, Y. (1989) J. Biol. Chem. 265, 1134]....

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00237a007

    authors： Lehrer SS,Stafford WF 3rd

    更新日期：1991-06-11 00:00:00

  • Pyruvate dehydrogenase kinase isoform 2 activity limited and further inhibited by slowing down the rate of dissociation of ADP.

    abstract：:Pyruvate dehydrogenase kinase 2 (PDK2) activity is enhanced by the dihydrolipoyl acetyltransferase core (E2 60mer) that binds PDK2 and a large number of its pyruvate dehydrogenase (E1) substrate. With E2-activated PDK2, K(+) at approximately 90 mM and Cl(-) at approximately 60 mM decreased the K(m) of PDK2 for ATP and...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi049488x

    authors： Bao H,Kasten SA,Yan X,Roche TE

    更新日期：2004-10-26 00:00:00

  • Function of arginine-234 and aspartic acid-271 in domain closure, cooperativity, and catalysis in Escherichia coli aspartate transcarbamylase.

    abstract：:Two mutant versions of Escherichia coli aspartate transcarbamylase were created by site-specific mutagenesis. Arg-234 of the 240s loop was replaced by serine in order to help deduce the function of the interactions that normally occur between Arg-234 and both Glu-50 and Gln-231 in the R state of the enzyme. The other ...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00423a022

    authors： Middleton SA,Kantrowitz ER

    更新日期：1988-11-15 00:00:00

  • Reversible unfolding of cytochrome c upon interaction with cardiolipin bilayers. 1. Evidence from deuterium NMR measurements.

    abstract：:Deuterium NMR has been used to investigate the structure and dynamic state of cytochrome c complexed with bilayers of cardiolipin. Reductive methylation was employed to prepare [N epsilon, N epsilon-C2H3]lysyl cytochrome c, and deuterium exchange provided labeling of backbone sites to give [amide-2H]cytochrome c or mo...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00230a010

    authors： Spooner PJ,Watts A

    更新日期：1991-04-23 00:00:00

  • Thioesterase portability and peptidyl carrier protein swapping in yersiniabactin synthetase from Yersinia pestis.

    abstract：:Multimodular enzymes, including polyketide synthases (PKSs), nonribosomal peptide synthetases (NRPSs), and mixed PKS/NRPS systems, contain functional domains with similar functions. Domain swapping and module fusion are potential powerful strategies for creating hybrid enzymes to synthesize modified natural products. ...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi047538s

    authors： Suo Z

    更新日期：2005-03-29 00:00:00

  • Activation of isolated NADH:ubiquinone reductase I (complex I) from Escherichia coli by detergent and phospholipids. Recovery of ubiquinone reductase activity and changes in EPR signals of iron-sulfur clusters.

    abstract：:NADH:ubiquinone oxidoreductase (NDH-1 or complex I) from Escherichia coli was purified using a combination of anion exchange chromatography and centrifugation in sucrose density gradient. The dependence of enzyme activity on detergent and phospholipids was studied. Artificial hexaammineruthenium reductase activity was...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi050134v

    authors： Sinegina L,Wikström M,Verkhovsky MI,Verkhovskaya ML

    更新日期：2005-06-14 00:00:00

  • Mechanism Underlying Anti-Markovnikov Addition in the Reaction of Pentalenene Synthase.

    abstract：:Most terpene synthase reactions follow Markovnikov rules for formation of high-energy carbenium ion intermediates. However, there are notable exceptions. For example, pentalenene synthase (PS) undergoes an initial anti-Markovnikov cyclization reaction followed by a 1,2-hydride shift to form an intermediate humulyl cat...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/acs.biochem.0c00518

    authors： Matos JO,Kumar RP,Ma AC,Patterson M,Krauss IJ,Oprian DD

    更新日期：2020-09-08 00:00:00

  • Crystal structure of homoserine transacetylase from Haemophilus influenzae reveals a new family of alpha/beta-hydrolases.

    abstract：:Homoserine transacetylase catalyzes one of the required steps in the biosynthesis of methionine in fungi and several bacteria. We have determined the crystal structure of homoserine transacetylase from Haemophilus influenzae to a resolution of 1.65 A. The structure identifies this enzyme to be a member of the alpha/be...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi051951y

    authors： Mirza IA,Nazi I,Korczynska M,Wright GD,Berghuis AM

    更新日期：2005-12-06 00:00:00

  • Membrane fusion and inverted phases.

    abstract：:We have found a correlation between liposome fusion kinetics and lipid phase behavior for several inverted phase forming lipids. N-Methylated dioleoylphosphatidylethanolamine (DOPE-Me), or mixtures of dioleoylphosphatidylethanolamine (DOPE) and dioleoylphosphatidylcholine (DOPC), will form an inverted hexagonal phase ...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00435a011

    authors： Ellens H,Siegel DP,Alford D,Yeagle PL,Boni L,Lis LJ,Quinn PJ,Bentz J

    更新日期：1989-05-02 00:00:00

  • Structural and functional modulation of the manganese cluster in Ca(2+)-depleted photosystem II induced by binding of the 24-kilodalton extrinsic protein.

    abstract：:Depletion of functional Ca2+ from photosystem (PS) II membranes impairs O2 evolution. Redox properties of the Mn cluster as probed by thermoluminescence were modified differently in Ca(2+)-depleted PSII depending on the procedure for Ca2+ extraction. Ca2+ depletion by low-pH treatment gave rise to an abnormally modifi...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00148a028

    authors： Ono T,Izawa S,Inoue Y

    更新日期：1992-08-25 00:00:00

  • The A-state of barnase.

    abstract：:The acid-induced denaturation of barnase and its mutants has been analyzed to search for partly-folded intermediates. Differential scanning calorimetry of barnase deviates from two-state behavior below pH 4.0 at low ionic strength, with the maximum discrepancy at pH 2.7. Addition of 200 mM KCl apparently restores the ...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00203a015

    authors： Sanz JM,Johnson CM,Fersht AR

    更新日期：1994-09-20 00:00:00

  • Characterization of the internal calcium(II) binding sites in dissolved insulin hexamer using europium(III) fluorescence.

    abstract：:The fluorescence of Eu(III) is used to study the nature of the Ca(II) binding sites in the central cavity of the two-zinc(II) insulin hexamer. The dependence of the Eu(III) fluorescence lifetime upon Eu(III) stoichiometry indicates that there are three identical Eu(III) binding sites present in the two-zinc(II) insuli...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00328a028

    authors： Alameda GK,Evelhoch JL,Sudmeier JL,Birge RR

    更新日期：1985-03-26 00:00:00

  • Microsecond folding of the cold shock protein measured by a pressure-jump technique.

    abstract：:A pressure-jump apparatus was employed in investigating the kinetics of protein unfolding and refolding. In the reaction cell, the pressure can be increased or decreased by 100-160 bar within 50-100 microseconds and then held constant. Thus, unfolding and refolding reactions in the time range from 70 microseconds to 7...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi982487i

    authors： Jacob M,Holtermann G,Perl D,Reinstein J,Schindler T,Geeves MA,Schmid FX

    更新日期：1999-03-09 00:00:00

  • 19F-NMR spin-spin relaxation (T2) method for characterizing volatile anesthetic binding to proteins. Analysis of isoflurane binding to serum albumin.

    abstract：:This paper characterizes the low-affinity ligand binding interactions of a fluorinated volatile anesthetic, isoflurane (CHF2OCHClCF3), with bovine serum albumin (BSA) using 19F-NMR transverse relaxation (T2). 19F-NMR spectra of isoflurane in aqueous BSA reveal a single isoflurane trifluoromethyl resonance, indicative ...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00146a007

    authors： Dubois BW,Evers AS

    更新日期：1992-08-11 00:00:00

  • Structure of pseudobactin A214, a siderophore from a bean-deleterious Pseudomonas.

    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

    authors： Buyer JS,Wright JM,Leong J

    更新日期：1986-09-23 00:00:00

  • The roles of active-site residues in the catalytic mechanism of trans-3-chloroacrylic acid dehalogenase: a kinetic, NMR, and mutational analysis.

    abstract：:trans-3-Chloroacrylic acid dehalogenase (CaaD) converts trans-3-chloroacrylic acid to malonate semialdehyde by the addition of H(2)O to the C-2, C-3 double bond, followed by the loss of HCl from the C-3 position. Sequence similarity between CaaD, an (alphabeta)(3) heterohexamer (molecular weight 47,547), and 4-oxalocr...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi030241u

    authors： Azurmendi HF,Wang SC,Massiah MA,Poelarends GJ,Whitman CP,Mildvan AS

    更新日期：2004-04-13 00:00:00

  • Methionine sulfoxide cytochrome c.

    abstract：:Cytochrome c has been chemically modified by methylene blue mediated photooxidation. It is established that the methionine residues of the protein have been specifically converted to methionine sulfoxide residues. No oxidation of any other amino acid residues or the cysteine thioether bridges of the molecule occurs du...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00650a029

    authors： Ivanetich KM,Bradshaw JJ,Kaminsky LS

    更新日期：1976-03-09 00:00:00

  • Investigation of an octapeptide inhibitor of Escherichia coli ribonucleotide reductase by transferred nuclear Overhauser effect spectroscopy.

    abstract：:Several peptides contained within the C-terminal sequence of the B2 subunit of Escherichia coli ribonucleotide reductase (RNR) were investigated for their ability to inhibit the enzyme, presumably by interfering with association of the B1 and B2 subunits. AcYLVGQIDSE, corresponding by sequence homology to a nonapeptid...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00247a008

    authors： Bushweller JH,Bartlett PA

    更新日期：1991-08-20 00:00:00

  • Gel to liquid-crystalline transition temperatures of water dispersions of two pairs of positional isomers of unsaturated mixed-acid phosphatidylcholines.

    abstract：:The gel to liquid-crystalline phase transition temperatures of dispersions of mixed-acid sn-1,2-lecithins which contain one unsaturated and one saturated fatty acid have been studied by differential scanning calorimetry. The temperature for 1-oleoyl-2-palmitoyl-sn-glycero-3-phosphocholine (containing no reversed isome...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00515a051

    authors： Davis PJ,Fleming BD,Coolbear KP,Keough KM

    更新日期：1981-06-09 00:00:00