Substitution of Tyr254 with Phe at the active site of flavocytochrome b2: consequences on catalysis of lactate dehydrogenation.

Abstract:

:A role for Tyr254 in L-lactate dehydrogenation catalyzed by flavocytochrome b2 has recently been proposed on the basis of the known active-site structure and of studies that had suggested a mechanism involving the initial formation of a lactate carbanion [Lederer, F., & Mathews, F.S. (1987) in Flavins and Flavoproteins, Proceedings of the Ninth International Symposium, Atlanta, GA, 1987 (Edmondson, D.E., & McCormick, D.B., Eds.) pp 133-142, Walter de Gruyter, Berlin]. This role is now examined after replacement of Tyr254 with phenylalanine. The kcat is decreased about 40-fold, Km for lactate appears unchanged, and the mainly rate-limiting step is still alpha-hydrogen abstraction, as judged from the steady-state deuterium isotope effect. Modeling studies with lactate introduced into the active site indicate two possible substrate conformations with different hydrogen-bonding partners for the substrate hydroxyl. If the hydrogen bond is formed with Tyr254, as was initially postulated, the mechanism must involve removal by His373 of the C2 hydrogen, with carbanion formation. If, in the absence of the Tyr254 phenol group, the hydrogen bond is formed with His373 N3, the substrate is positioned in such a way that the reaction must proceed by hydride transfer. Therefore the mechanism of the Y254F enzyme was investigated so as to distinguish between the two mechanistic possibilities. 2-Hydroxy-3-butynoate behaves with the mutant as a suicide reagent, as with the wild-type enzyme. Similarly, the mutant protein also catalyzes the reduction and the dehydrohalogenation of bromopyruvate under transhydrogenation conditions.(ABSTRACT TRUNCATED AT 250 WORDS)

journal_name

Biochemistry

journal_title

Biochemistry

authors

Dubois J,Chapman SK,Mathews FS,Reid GA,Lederer F

doi

10.1021/bi00479a008

subject

Has Abstract

pub_date

1990-07-10 00:00:00

pages

6393-400

issue

27

eissn

0006-2960

issn

1520-4995

journal_volume

29

pub_type

杂志文章
  • Tandem GA residues on opposite sides of the loop in molecular beacon-like DNA hairpins compact the loop and increase hairpin stability.

    abstract::The free solution electrophoretic mobilities and thermal stabilities of hairpins formed by two complementary 26-nucleotide oligomers have been measured by capillary electrophoresis. The oligomers are predicted to form molecular beacon-like hairpins with 5 bp stems and 16 nucleotides in the loop. One hairpin, called ha...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi201263n

    authors: Chang CY,Stellwagen NC

    更新日期:2011-10-25 00:00:00

  • Bleaching kinetics of artificial visual pigments with modifications near the ring-polyene chain connection.

    abstract::Absorbance difference spectra were recorded at 20 degrees C from 30 ns to milliseconds after photolysis of lauryl maltoside suspensions of artificial visual pigments derived from 9-cis isomers of 5-ethylretinal, 8,16-methanoretinal (a 6-s-trans-bicyclic analogue), or 5-demethyl-8-methylretinal. In all three pigments, ...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi011461f

    authors: Szundi I,de Lera AR,Pazos Y,Alvarez R,Oliana M,Sheves M,Lewis JW,Kliger DS

    更新日期:2002-02-12 00:00:00

  • Diepoxybutane cross-links DNA at 5'-GNC sequences.

    abstract::Epoxides are cancer-causing agents chemically analogous to the nitrogen mustards, a family of powerful antitumor drugs. We found that the DNA interstrand cross-linking sequence preference of diepoxybutane is the same as that of the mustard mechlorethamine: 5'-GNC. Therefore, the genomic site of cross-linking alone can...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00059a034

    authors: Millard JT,White MM

    更新日期:1993-03-02 00:00:00

  • Mechanism and role of cooperative binding of bacteriophage fd gene 5 protein to single-stranded deoxyribonucleic acid.

    abstract::The highly cooperative binding of fd gene 5 to single-stranded DNA was studied kinetically by rapid photo-cross-linking and stopped-flow UV absorption measurements. The observed change in absorbance was shown to be due to the binding by direct evidence of rapid photo-cross-linking of the bound proteins to fd DNA. The ...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00294a028

    authors: Shimamoto N,Utiyama H

    更新日期:1983-12-06 00:00:00

  • Chromomycin dimer-DNA oligomer complexes. Sequence selectivity and divalent cation specificity.

    abstract::This paper reports on a solution NMR characterization of the sequence selectivity and metal ion specificity in chromomycin-DNA oligomer complexes in the presence of divalent cations. The sequence selectivity studies have focused on chromomycin complexes with the self-complementary d(A1-A2-G3-G4-C5-C6-T7-T8) duplex con...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00501a012

    authors: Gao XL,Patel DJ

    更新日期:1990-12-11 00:00:00

  • Binding of calcium to anticoagulant protein S: role of the fourth EGF module.

    abstract::Protein S is an anticoagulant protein containing a Gla (enclosing gamma-carboxyglutamic acids) module, a TSR (thrombin sensitive region) module, four EGF (epidermal growth factor)-like modules, and a SHBG (sex hormone binding globulin)-like region. Protein S is a cofactor to activated protein C (APC) in the degradatio...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi0601151

    authors: Persson KE,Stenflo J,Linse S,Stenberg Y,Preston RJ,Lane DA,Rezende SM

    更新日期:2006-09-05 00:00:00

  • The crystal structure of human procathepsin K.

    abstract::Cathepsin K is a cysteine protease present in human osteoclasts that plays an important role in bone resorption. Cathepsin K is synthesized as an inactive proenzyme and activated under conditions of low pH. Autoproteolytic processing of the N-terminal 99 amino acid propeptide produces the active, mature form of cathep...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi9822271

    authors: LaLonde JM,Zhao B,Janson CA,D'Alessio KJ,McQueney MS,Orsini MJ,Debouck CM,Smith WW

    更新日期:1999-01-19 00:00:00

  • Poly(dA).poly(dT) exists in an unusual conformation under physiological conditions: propidium binding to poly(dA).poly(dT) and poly[d(A-T)].poly[d(A-T)].

    abstract::The binding of propidium to poly(dA).poly(dT) [poly(dA.dT)] and to poly[d(A-T)].poly[d(A-T)] [poly[d(A-T)2]] has been compared under a variety of solution conditions by viscometric titrations, binding studies, and kinetic experiments. The binding of propidium to poly[d(A-T)2] is quite similar to its binding to calf th...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00336a029

    authors: Wilson WD,Wang YH,Krishnamoorthy CR,Smith JC

    更新日期:1985-07-16 00:00:00

  • Identification of polyphosphate recognition sites communicating with actin sites on the skeletal myosin subfragment 1 heavy chain.

    abstract::Using the thrombin-cut [68-30 kilodalton (kDa)] myosin subfragment 1 (S-1) whose heavy chain has been selectively split within the central 50-kDa region, at Lys-560, with concomitant specific alterations of the ATPase and actin binding properties [Chaussepied, P., Mornet, D., Audemard, E., Derancourt, J., & Kassab, R....

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00369a013

    authors: Chaussepied P,Mornet D,Kassab R

    更新日期:1986-10-21 00:00:00

  • Thermotropic and mixing behavior of mixed-chain phosphatidylcholines with molecular weights identical with that of L-alpha-dipalmitoylphosphatidylcholine.

    abstract::The thermotropic phase behavior of 10 mixed-chain phosphatidylcholines, in excess water, has been examined and compared with that of identical-chain C(16):C(16)PC by using high-resolution differential scanning calorimetry (DSC). The molecular weights (MW) of these 11 molecular species are the same, but their delta C/C...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00243a022

    authors: Bultmann T,Lin HN,Wang ZQ,Huang CH

    更新日期:1991-07-23 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

  • Location and interactions of phospholipid and cholesterol in human low density lipoprotein from 31P nuclear magnetic resonance.

    abstract::The major phospholipids, phosphatidylcholine and spingomyelin, of low density lipoprotein (LDL) are accessible to small amounts of Pr3+, suggesting that the head groups of all mobile phospholipids are on the surface of the particle in contact with the aqueous medium. The major source of the nuclear Overhauser effect e...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00607a002

    authors: Yeagle PL,Martin RB,Pottenger L,Langdon RG

    更新日期:1978-07-11 00:00:00

  • The complete amino acid sequence of the major component myoglobin of dwarf sperm whale (Kogia simus).

    abstract::The complete amino acid sequence of the major component myoglobin from the dwarf sperm whale, Kogia simus, was determined by specific cleavage of the protein to obtain large peptides which are readily degraded by the automatic sequenator. Three easily separable peptides were obtained by cleaving the protein at its two...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00624a010

    authors: Dwulet FE,Jones BN,Lehman LD,Gurd FR

    更新日期:1977-03-08 00:00:00

  • Formation of a porphyrin pi-cation radical in the fluoride complex of horseradish peroxidase.

    abstract::Horseradish peroxidase (HRP) was oxidized by IrCl6(2-) to a mixture of compounds I and II, the rate of oxidation and the ratio of the mixture being greatly affected by pH (Hayashi & Yamazaki, 1979). Oxidation of HRP by IrCl6(2-) in the presence of fluoride was significantly accelerated. This resulted in the formation ...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00046a003

    authors: Farhangrazi ZS,Sinclair R,Powers L,Yamazaki I

    更新日期:1995-11-21 00:00:00

  • Nucleotide clusters in deoxyribonucleic acids: sequence analysis of DNA using pyrimidine oligonucleotides as primers in the DNA polymerase I repair reaction.

    abstract::Pyrimidine oligonucleotides have been shown to prime the E. coli DNA polymerase I repair reaction, specifically and reproducibly. DNA molecules up to 30 nucleotides long have been obtained from the extension of oligopyrimidine primers, 9 to 11 nucleotides long isolated from the complementary (minus) strand of bacterio...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00598a015

    authors: Kaptein JS,Spencer JH

    更新日期:1978-03-07 00:00:00

  • Order and dynamics in mixtures of membrane glucolipids from Acholeplasma laidlawii studied by 2H NMR.

    abstract::The two dominant glucolipids in Acholeplasma laidlawii, viz., 1,2-diacyl-3-O-(alpha-D-glucopyranosyl)-sn-glycerol (MGlcDG) and 1,2-diacyl-3-O-[alpha-D-glucopyranosyl-(1----2)-O-alpha-D-glucopyranosyl ]- sn-glycerol (DGlcDG), have markedly different phase behavior. MGlcDG has an ability to form nonlamellar phases, wher...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00234a012

    authors: Eriksson PO,Rilfors L,Wieslander A,Lundberg A,Lindblom G

    更新日期:1991-05-21 00:00:00

  • Purification and partial amino acid sequence of papain-solubilized class II transplantation antigens.

    abstract::Papain-solubilized human class II (HLA-DR) antigens have been purified from cadaveric spleens by ion-exchange chromatography, gel chromatography, and immunosorbent purification. The isolated papain-solubilized antigens comprised two subunits with apparent molecular weights of 23 000 and 30 000, respectively. The circu...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00264a036

    authors: Wiman K,Claesson L,Rask L,Trägårdh L,Peterson PA

    更新日期:1982-10-12 00:00:00

  • Differential susceptibility of transferrin glycoforms to chymotrypsin: a proteomics approach to the detection of carbohydrate-deficient transferrin.

    abstract::Transferrin exhibits heterogeneity in glycosylation characteristic of pathological changes in alcohol abuse and congenital disorders in glycosylation. This study investigated an alternative approach in the detection of carbohydrate-deficient transferrin based on the premise that glycosylation may afford some degree of...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi051749v

    authors: Valmu L,Kalkkinen N,Husa A,Rye PD

    更新日期:2005-12-13 00:00:00

  • Aluminum fluoride activation of bovine transducin induces two distinct conformational changes in the alpha subunit.

    abstract::We have used resonance energy transfer to read out the interactions of the alpha subunit of transducin (alpha T) with the transducin beta gamma subunit complex (beta gamma T) and to compare the rate of aluminum fluoride-induced alpha T activation, as reflected by the enhancement of the alpha T tryptophan fluorescence,...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00199a046

    authors: Mittal R,Cerione RA,Erickson JW

    更新日期:1994-08-23 00:00:00

  • Mutant IDH1 enhances the production of 2-hydroxyglutarate due to its kinetic mechanism.

    abstract::The human, cytosolic enzyme isocitrate dehydrogenase 1 (IDH1) reversibly converts isocitrate to α-ketoglutarate (αKG). Cancer-associated somatic mutations in IDH1 result in a loss of this normal function but a gain in a new or neomorphic ability to convert αKG to the oncometabolite 2-hydroxyglutarate (2HG). To improve...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi400514k

    authors: Rendina AR,Pietrak B,Smallwood A,Zhao H,Qi H,Quinn C,Adams ND,Concha N,Duraiswami C,Thrall SH,Sweitzer S,Schwartz B

    更新日期:2013-07-02 00:00:00

  • The multifunctional protein nucleophosmin (NPM1) is a human linker histone H1 chaperone.

    abstract::Linker histone H1 plays an essential role in chromatin organization. Proper deposition of linker histone H1 as well as its removal is essential for chromatin dynamics and function. Linker histone chaperones perform this important task during chromatin assembly and other DNA-templated phenomena in the cell. Our in vitr...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi101835j

    authors: Gadad SS,Senapati P,Syed SH,Rajan RE,Shandilya J,Swaminathan V,Chatterjee S,Colombo E,Dimitrov S,Pelicci PG,Ranga U,Kundu TK

    更新日期:2011-04-12 00:00:00

  • Posttranslational Chemical Mutagenesis: To Reveal the Role of Noncatalytic Cysteine Residues in Pathogenic Bacterial Phosphatases.

    abstract::The field of chemical site-selective modification of proteins has progressed extensively in recent decades to enable protein functionalization for imaging, drug delivery, and functional studies. In this Perspective, we provide detailed insight into an alternative use of site-selective protein chemistry to probe the ro...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/acs.biochem.8b00639

    authors: Bertoldo JB,Terenzi H,Hüttelmaier S,Bernardes GJL

    更新日期:2018-10-30 00:00:00

  • Normal modes of prion proteins: from native to infectious particle.

    abstract::Prion proteins (PrP) are the infectious agent in transmissible spongiform encephalopathies (i.e., mad cow disease). To be infectious, prion proteins must undergo a conformational change involving a decrease in α-helical content along with an increase in β-strand content. This conformational change was evaluated by mea...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi1010514

    authors: Samson AO,Levitt M

    更新日期:2011-03-29 00:00:00

  • Ruthenium-iron hybrid hemoglobins as a model for partially liganded hemoglobin: oxygen equilibrium curves and resonance Raman spectra.

    abstract::The structure and function of iron(II)-ruthenium(II) hybrid hemoglobins alpha(Ru-CO)2 beta(Fe)2 and alpha(Fe)2 beta(Ru-CO)2, which can serve as models for the intermediate species of the oxygenation step in native human adult hemoglobin, were investigated by measuring oxygen equilibrium curves and the Fe(II)-N epsilon...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00447a050

    authors: Ishimori K,Tsuneshige A,Imai K,Morishima I

    更新日期:1989-10-17 00:00:00

  • Deletion of exon 21 of the insulin receptor eliminates tyrosine kinase activity but preserves mitogenic signaling.

    abstract::To study the function of exon 21 of the insulin receptor, a mutant human insulin receptor lacking this domain was constructed. The mutant HIR delta E21 cDNA was transfected into Rat-1 fibroblasts and stable cell lines were selected. The HIR delta E21 receptors were expressed on the cell surface, and they bound insulin...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00212a021

    authors: Rolband GC,Williams JF,Webster NJ,Hsu D,Olefsky JM

    更新日期:1993-12-14 00:00:00

  • The Inflection Point Hypothesis: The Relationship between the Temperature Dependence of Enzyme-Catalyzed Reaction Rates and Microbial Growth Rates.

    abstract::The temperature dependence of biological rates at different scales (from individual enzymes to isolated organisms to ecosystem processes such as soil respiration and photosynthesis) is the subject of much historical and contemporary research. The precise relationship between the temperature dependence of enzyme rates ...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/acs.biochem.0c00530

    authors: Prentice EJ,Hicks J,Ballerstedt H,Blank LM,Liáng LNL,Schipper LA,Arcus VL

    更新日期:2020-09-29 00:00:00

  • Labeling of two different regions of the nucleotide binding site of the uncoupling protein from brown adipose tissue mitochondria with two ATP analogs.

    abstract::The nucleotide binding site of the uncoupling protein (UCP) from brown adipose tissue was mapped by photoaffinity labeling with 2-azidoadenosine 5'-triphosphate (2-azido-ATP) and by affinity labeling with 3'-O-(5-fluoro-2,4-dinitrophenyl)adenosine 5'-triphosphate (FDNP-ATP). Both analogs bind with high affinity and sp...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00158a017

    authors: Mayinger P,Klingenberg M

    更新日期:1992-11-03 00:00:00

  • Water-mediated substrate/product discrimination: the product complex of thymidylate synthase at 1.83 A.

    abstract::In an irreversible enzyme-catalyzed reaction, strong binding of the products would lead to substantial product inhibition. The X-ray crystal structure of the product complex of thymidylate synthase (1.83-A resolution, R factor = 0.183 for all data between 7.0 and 1.83 A) identifies a bound water molecule that serves t...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00172a029

    authors: Fauman EB,Rutenber EE,Maley GF,Maley F,Stroud RM

    更新日期:1994-02-15 00:00:00

  • Model for general acid-base catalysis by the hammerhead ribozyme: pH-activity relationships of G8 and G12 variants at the putative active site.

    abstract::We have used nucleobase substitution and kinetic analysis to test the hypothesis that hammerhead catalysis occurs by a general acid-base mechanism, in which nucleobases are directly involved in deprotonation of the attacking 2'-hydroxyl group and protonation of the 5'-oxygen that serves as the leaving group in the cle...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi047941z

    authors: Han J,Burke JM

    更新日期:2005-05-31 00:00:00

  • Effects of phosphorothioate and 2-amino groups in hammerhead ribozymes on cleavage rates and Mg2+ binding.

    abstract::Mg2+ is important for the RNase activity of the hammerhead ribozyme. To investigate the binding properties of Mg2+ to the hammerhead ribozyme, cleavage rates and CD spectra for substrates containing inosine or guanosine at the cleavage site were measured. The 2-amino group of this guanosine interfered with the rate of...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00235a005

    authors: Koizumi M,Ohtsuka E

    更新日期:1991-05-28 00:00:00