Entropy effects on protein hinges: the reaction catalyzed by triosephosphate isomerase.

Abstract:

:Many proteins utilize segmental motions to catalyze a specific reaction. The Omega loop of triosephosphate isomerase (TIM) is important for preventing the loss of the reactive enediol(ate) intermediate. The loop opens and closes even in the absence of the ligand, and the loop itself does not change conformation during movement. The conformational changes are localized to two hinges at the loop termini. Glycine is never observed in native TIM hinge sequences. In this paper, the hypothesis that limited access to conformational space is a requirement for protein hinges involved in catalysis was tested. The N-terminal hinge was mutated to P166/V167G/W168G (PGG), and the C-terminal hinge was mutated to K174G/T175G/A176G (GGG) in chicken TIM. The single-hinge mutants PGG and GGG had k(cat) values 200-fold lower than that of the wild type and K(m) values 10-fold higher. The k(cat) of double-hinge mutant P166/V167G/W168G/K174G/T175G/A176G was reduced 2500-fold; the K(m) was 10-fold higher. A combination of primary kinetic isotope effect measurements, isothermal calorimetric measurements, and (31)P NMR spectroscopic titration with the inhibitor 2-phosphoglycolate revealed that the mutants have a different ligand-binding mode than that of the wild-type enzyme. The predominant conformations of the mutants even in the presence of the inhibitor are loop-open conformations. In conclusion, mutation of the hinge residues to glycine resulted in the sampling of many more hinge conformations with the consequence that the population of the active-closed conformation is reduced. This reduced population results in a reduced catalytic activity.

journal_name

Biochemistry

journal_title

Biochemistry

authors

Xiang J,Jung JY,Sampson NS

doi

10.1021/bi049208d

subject

Has Abstract

pub_date

2004-09-14 00:00:00

pages

11436-45

issue

36

eissn

0006-2960

issn

1520-4995

journal_volume

43

pub_type

杂志文章
  • Protein arginine deiminase 4: evidence for a reverse protonation mechanism.

    abstract::The presumed role of an overactive protein arginine deiminase 4 (PAD4) in the pathophysiology of rheumatoid arthritis (RA) suggests that PAD4 inhibitors could be used to treat an underlying cause of RA, potentially offering a mechanism to stop further disease progression. Thus, the development of such inhibitors is of...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi700095s

    authors: Knuckley B,Bhatia M,Thompson PR

    更新日期:2007-06-05 00:00:00

  • Mechanism of interactions of alpha-naphthoflavone with cytochrome P450 3A4 explored with an engineered enzyme bearing a fluorescent probe.

    abstract::Design of a partially cysteine-depleted C98S/C239S/C377S/C468A cytochrome P450 3A4 mutant designated CYP3A4(C58,C64) allowed site-directed incorporation of thiol-reactive fluorescent probes into alpha-helix A. The site of modification was identified as Cys-64 with the help of CYP3A4(C58) and CYP3A4(C64), each bearing ...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi061944p

    authors: Tsalkova TN,Davydova NY,Halpert JR,Davydov DR

    更新日期:2007-01-09 00:00:00

  • Structural and functional effects of tryptophans inserted into the membrane-binding and substrate-binding sites of human group IIA phospholipase A2.

    abstract::Phospholipase A(2) (PLA(2)) enzymes become activated by binding to biological membranes and hydrolyze phospholipids to free fatty acids and lyso-phospholipids, the precursors of inflammatory mediators. To understand the functional significance of amino acid residues at key positions, we have studied the effects of the...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi061440r

    authors: Nemec KN,Pande AH,Qin S,Bieber Urbauer RJ,Tan S,Moe D,Tatulian SA

    更新日期:2006-10-17 00:00:00

  • Identification of 9-cis,13-cis-retinoic acid as a major circulating retinoid in plasma.

    abstract::In the course of measuring the concentrations of retinoic acids (RA) in bovine plasma, a major peak was observed which comigrated with 9-cis-RA on normal-phase high-performance liquid chromatography. Rechromatography of this retinoic on reverse-phase high-performance liquid chromatography showed that it was distinct f...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00004a013

    authors: Horst RL,Reinhardt TA,Goff JP,Nonnecke BJ,Gambhir VK,Fiorella PD,Napoli JL

    更新日期:1995-01-31 00:00:00

  • Endogenous 5-methylcytosine protects neighboring guanines from N7 and O6-methylation and O6-pyridyloxobutylation by the tobacco carcinogen 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone.

    abstract::All CG dinucleotides along exons 5-8 of the p53 tumor suppressor gene contain endogenous 5-methylcytosine (MeC). These same sites (e.g., codons 157, 158, 245, 248, and 273) are mutational hot spots in smoking-induced lung cancer. Several groups used the UvrABC endonuclease incision assay to demonstrate that methylated...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi035259j

    authors: Ziegel R,Shallop A,Upadhyaya P,Jones R,Tretyakova N

    更新日期:2004-01-20 00:00:00

  • Protein Synthesis with Ribosomes Selected for the Incorporation of β-Amino Acids.

    abstract::In an earlier study, β³-puromycin was used for the selection of modified ribosomes, which were utilized for the incorporation of five different β-amino acids into Escherichia coli dihydrofolate reductase (DHFR). The selected ribosomes were able to incorporate structurally disparate β-amino acids into DHFR, in spite of...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/acs.biochem.5b00389

    authors: Maini R,Chowdhury SR,Dedkova LM,Roy B,Daskalova SM,Paul R,Chen S,Hecht SM

    更新日期:2015-06-16 00:00:00

  • Design of a selective insulin receptor tyrosine kinase inhibitor and its effect on glucose uptake and metabolism in intact cells.

    abstract::An inhibitor of the insulin receptor tyrosine kinase (IRTK), (hydroxy-2-naphthalenyl-methyl) phosphonic acid, was designed and synthesized and was shown to be an inhibitor of the biological effects of insulin in vitro. With a wheat germ purified human placental insulin receptor preparation, this compound inhibited the...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00439a053

    authors: Saperstein R,Vicario PP,Strout HV,Brady E,Slater EE,Greenlee WJ,Ondeyka DL,Patchett AA,Hangauer DG

    更新日期:1989-06-27 00:00:00

  • Enzymatic properties of proteolytic derivatives of human alpha-thrombin.

    abstract::The use of derivatives of alpha-thrombin obtained by limited proteolysis, that have only a single peptide bond cleaved, allowed the unequivocal correlation between the change in covalent structure and alteration of the enzymatic properties. beta T-Thrombin contains a single cleavage in the surface loop corresponding t...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00406a049

    authors: Hofsteenge J,Braun PJ,Stone SR

    更新日期:1988-03-22 00:00:00

  • Mutation of Arg-166 of alkaline phosphatase alters the thio effect but not the transition state for phosphoryl transfer. Implications for the interpretation of thio effects in reactions of phosphatases.

    abstract::It has been suggested that the mechanism of alkaline phosphatase (AP) is associative, or triester-like, because phosphorothioate monoesters are hydrolyzed by AP approximately 10(2)-fold slower than phosphate monoesters. This "thio effect" is similar to that observed for the nonenzymatic hydrolysis of phosphate trieste...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi000899x

    authors: Holtz KM,Catrina IE,Hengge AC,Kantrowitz ER

    更新日期:2000-08-08 00:00:00

  • Site-directed sulfhydryl labeling of the lactose permease of Escherichia coli: helices IV and V that contain the major determinants for substrate binding.

    abstract::Helices IV and V in the lactose permease of Escherichia coli contain the major determinants for substrate binding [Glu126 (helix IV), Arg144 (helix V), and Cys148 (helix V)]. Structural and dynamic features of this region were studied by using site-directed sulfhydryl modification of 48 single-Cys replacement mutants ...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi010866x

    authors: Kwaw I,Zen KC,Hu Y,Kaback HR

    更新日期:2001-09-04 00:00:00

  • Identification of human CYP2C19 residues that confer S-mephenytoin 4'-hydroxylation activity to CYP2C9.

    abstract::CYP2C19 is selective for the 4'-hydroxylation of S-mephenytoin while the highly similar CYP2C9 has little activity toward this substrate. To identify critical amino acids determining the specificity of human CYP2C19 for S-mephenytoin 4'-hydroxylation, we constructed chimeras by replacing portions of CYP2C9 containing ...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi001678u

    authors: Tsao CC,Wester MR,Ghanayem B,Coulter SJ,Chanas B,Johnson EF,Goldstein JA

    更新日期:2001-02-20 00:00:00

  • Pre-steady-state kinetics reveal a slow isomerization of the enzyme-NAD complex in the NAD-malic enzyme reaction.

    abstract::Stopped-flow experiments obtained in the pre-steady-state time scale of the NAD-malic enzyme reaction exhibit a lag prior to the attainment of steady state. Previous results from isotope effect studies in which the deuterium isotope effect on Vmax decreases to a value of 1 at low pH have been interpreted as suggesting...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00059a007

    authors: Rajapaksa R,Abu-Soud H,Raushel FM,Harris BG,Cook PF

    更新日期:1993-03-02 00:00:00

  • Conformational equilibrium of an enzyme catalytic site in the allosteric transition.

    abstract::The dynamic equilibrium of a catalytic site between active and inactive conformations, the missing link between the structure and function of allosteric enzymes, was identified using protein engineering and NMR techniques. Kinetic analyses of the wild-type and three mutants of Thermus L-lactate dehydrogenase establish...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00138a017

    authors: Koide S,Yokoyama S,Matsuzawa H,Miyazawa T,Ohta T

    更新日期:1992-06-16 00:00:00

  • Identification and characterization of Mg2+ binding sites in isolated alpha and beta subunits of H(+)-ATPase from Bacillus PS3.

    abstract::The properties of the nucleotide binding sites in the isolated beta and alpha subunits of H(+)-ATPase from Bacillus PS3 (TF1) have been examined by studying the EPR properties of bound VO(2+), which is a paramagnetic probe for the native Mg2+ cation cofactor. The amino acid ligands of the VO2+ complexes with the isola...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi9910732

    authors: Zimmermann JL,Amano T,Sigalat C

    更新日期:1999-11-16 00:00:00

  • Roles of aquaporin-4 isoforms and amino acids in square array assembly.

    abstract::Aquaporin-4 (AQP4) is a water channel found at high concentrations around blood vessels in the brain and is organized into elaborate assemblies called square arrays. The natural functions of AQP4 and the square arrays remain unknown, but under pathophysiological conditions, AQP4 has been shown to influence brain edema...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi802231q

    authors: Strand L,Moe SE,Solbu TT,Vaadal M,Holen T

    更新日期:2009-06-30 00:00:00

  • Structure of the human pancreatic cholesterol esterase gene.

    abstract::The gene for human pancreatic cholesterol esterase consists of 11 exons and 10 introns and is 9.2 kb in length. The last and longest exon (841 nucleotides) is unique to the human gene. Functional amino acids are encoded on separate exons. The leader sequence is encoded by a single exon which carries two additional N-t...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00141a017

    authors: Kumar BV,Aleman-Gomez JA,Colwell N,Lopez-Candales A,Bosner MS,Spilburg CA,Lowe M,Lange LG

    更新日期:1992-07-07 00:00:00

  • Site specific combinations of stabilizing and destabilizing amino acid replacements in yeast cytochrome c: in vivo and in vitro effects.

    abstract::Oligonucleotide-directed mutagenesis in vivo was used to create destabilizing site specific changes at position 6 in iso-1-cytochrome c in Saccharomyces cerevisiae. These changes were made in combination with the stabilizing N52I substitution. The resulting proteins showed that a variety of forces and interactions are...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00021a023

    authors: Linske-O'Connell LI,Sherman F,McLendon G

    更新日期:1995-05-30 00:00:00

  • A Modified ToxT Inhibitor Reduces Vibrio cholerae Virulence in Vivo.

    abstract::We have previously designed and synthesized small-molecule inhibitors that reduce Vibrio cholerae virulence in vitro by targeting the transcription factor ToxT. Here we report the synthesis and biological activity of derivatives of our previous bicyclic, fatty acid-like inhibitors. All of the synthesized derivatives s...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/acs.biochem.8b00667

    authors: Woodbrey AK,Onyango EO,Kovacikova G,Kull FJ,Gribble GW

    更新日期:2018-09-25 00:00:00

  • Covalent structure of collagen: amino acid sequence of chick skin collagen alpha1(1)-CB6B.

    abstract::The amino acid sequence of the peptide alpha1(I)-CB6B, the carboxyl-terminal peptide containing 91 residues of alpha1(I) chain of chick skin collagen, is described. The amino acid sequence was determined by automated Edman degradation of intact peptide plus its tryptic and thermolytic peptides. The comparison of this ...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00619a018

    authors: Dixit SN,Seyer JM,Kang AH,Gross J

    更新日期:1978-12-26 00:00:00

  • Phosphoinositide-specific phospholipase C-delta 1: effect of monolayer surface pressure and electrostatic surface potentials on activity.

    abstract::We added phospholipase C-delta 1 (PLC-delta) to the aqueous subphase beneath monolayers formed from mixtures of phosphatidylinositol 4,5-bisphosphate (2% PIP2), phosphatidylserine (33% PS), and phosphatidylcholine (65% PC) and then measured the initial rate of hydrolysis of PIP2 after addition of 10 microM free calciu...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00166a006

    authors: Rebecchi M,Boguslavsky V,Boguslavsky L,McLaughlin S

    更新日期:1992-12-29 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

  • 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

  • Computational Study of Binding of μ-Conotoxin GIIIA to Bacterial Sodium Channels NaVAb and NaVRh.

    abstract::Structures of several voltage-gated sodium (NaV) channels from bacteria have been determined recently, but the same feat might not be achieved for the mammalian counterparts in the near future. Thus, at present, computational studies of the mammalian NaV channels have to be performed using homology models based on the...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/acs.biochem.5b01324

    authors: Patel D,Mahdavi S,Kuyucak S

    更新日期:2016-03-29 00:00:00

  • N-linked neutral sugar chains of aminopeptidase N purified from rat small intestinal brush-border membrane.

    abstract::Neutral oligosaccharides, which accounted for 74% of the total N-linked sugar chains released by hydrazinolysis of rat small intestinal aminopeptidase N, were investigated on a structural basis. They are mainly composed of complex-type sugar chains with tri- and tetraantennary structures, and small amounts of high man...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00101a028

    authors: Takasaki S,Erickson RH,Kim YS,Kochibe N,Kobata A

    更新日期:1991-09-17 00:00:00

  • Phosphonate analogues of carboxypeptidase A substrates are potent transition-state analogue inhibitors.

    abstract::Analogues of tri- and tetrapeptide substrates of carboxypeptidase A in which the scissile peptide linkage is replaced with a phosphonate moiety (-PO2--O-) were synthesized and evaluated as inhibitors of the enzyme. The inhibitors terminated with either L-lactate or L-phenyllactate [designated (O) Ala and (O) Phe, resp...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00441a022

    authors: Hanson JE,Kaplan AP,Bartlett PA

    更新日期:1989-07-25 00:00:00

  • Human SAS-6 C-Terminus Nucleates and Promotes Microtubule Assembly in Vitro by Binding to Microtubules.

    abstract::Centrioles are essential components of the animal centrosome and play crucial roles in the formation of cilia and flagella. They are cylindrical structures composed of nine triplet microtubules organized around a central cartwheel. Recent studies have identified spindle assembly abnormal protein SAS-6 as a critical co...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/acs.biochem.5b00978

    authors: Gupta H,Badarudeen B,George A,Thomas GE,Gireesh KK,Manna TK

    更新日期:2015-10-20 00:00:00

  • Contribution of basic residues of the 70-80-loop to heparin binding and anticoagulant function of activated protein C.

    abstract::The role of basic residues of the 70-80-loop, Arg(74), Arg(75), and Lys(78) (chymotrypsin numbering) in the catalytic function of activated protein C (APC) was investigated by expressing mutants of protein C in which these residues were replaced with Ala in three separate constructs. Following purification to homogene...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi015899r

    authors: Yang L,Manithody C,Rezaie AR

    更新日期:2002-05-14 00:00:00

  • Effect of subunit III removal on control of cytochrome c oxidase activity by pH.

    abstract::Studies were undertaken to assess the postulated involvement of subunit III in the proton-linked functions of cytochrome c oxidase. The effect of pH on the steady-state kinetic [corrected] parameters of subunit III containing and subunit III depleted cytochrome oxidase was determined by using beef heart and rat liver ...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00417a016

    authors: Gregory LC,Ferguson-Miller S

    更新日期:1988-08-23 00:00:00

  • Targeted generation of DNA strand breaks using pyrene-conjugated triplex-forming oligonucleotides.

    abstract::Gene targeting by triplex-forming oligonucleotides (TFOs) has proven useful for gene modulation in vivo. Photoreactive molecules have been conjugated to TFOs to direct sequence-specific damage in double-stranded DNA. However, the photoproducts are often repaired efficiently in cells. This limitation has led to the sea...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi7024029

    authors: Benfield AP,Macleod MC,Liu Y,Wu Q,Wensel TG,Vasquez KM

    更新日期:2008-06-10 00:00:00

  • Utilization of Substrate Intrinsic Binding Energy for Conformational Change and Catalytic Function in Phosphoenolpyruvate Carboxykinase.

    abstract::Phosphoenolpyruvate carboxykinase (PEPCK) is an essential metabolic enzyme operating in the gluconeogenesis and glyceroneogenesis pathways. Previous work has demonstrated that the enzyme cycles between a catalytically inactive open state and a catalytically active closed state. The transition of the enzyme between the...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/acs.biochem.5b01215

    authors: Johnson TA,Mcleod MJ,Holyoak T

    更新日期:2016-01-26 00:00:00