当前位置： SCI文献检索 > BIOCHEMISTRY期刊下所有文献 > Stopped-flow cryoenzymological investigation of the pre-steady-state kinetics of hydrolysis of Leu-Gly-NHNH-Dns by leucine aminopeptidase.

Stopped-flow cryoenzymological investigation of the pre-steady-state kinetics of hydrolysis of Leu-Gly-NHNH-Dns by leucine aminopeptidase.

Abstract：

:Stopped-flow fluorescence experiments have been carried out to study the steady-state kinetics of hydrolysis of Leu-Gly-NHNH-Dns [Dns = 5-(dimethylamino)naphthalene-1-sulfonyl] by porcine kidney cytosol leucine aminopeptidase (LAP) in 50% v/v methanol/buffer solution at ambient temperature and the pre-steady-state kinetics of this reaction in the -35 to 0 degrees C temperature range. Experiments have been carried out on LAP species containing Mg(II), Mn(II), Cu(II), Ni(II), Zn(II), and no metal ion at the regulatory metal binding site. At ambient temperatures, the stopped-flow fluorescence changes observed on hydrolysis of the substrate have been used to measure the steady-state kinetic parameters kcat and KM. The results show that 50% v/v methanol lowers the values of kcat from 2- to 12-fold compared to the reactions in the absence of methanol for all of the metallo-LAP, but that the values of KM are essentially unaffected. The pre-steady-state reactions carried out under nonturnover conditions at -35 degrees C reveal a new relaxation for LAP species with Ni(II), Cu(II), and Zn(II) in the regulatory site. The value of kobsd for this relaxation reaches a plateau at high substrate concentrations, and the magnitude of its fluorescence change at a fixed concentration of substrate is proportional to the enzyme concentration. Thus, this relaxation corresponds to the production and decay of a new enzyme-substrate intermediate not observed at higher temperatures whose fluorescence differs from that of the succeeding intermediate that is normally seen above -26 degrees C.

journal_name

Biochemistry

journal_title

Biochemistry

authors

Lin WY,Lin SH,Morris RJ,Van Wart HE

doi

10.1021/bi00414a018

subject

Has Abstract

pub_date

1988-07-12 00:00:00

pages

5068-74

issue

14

eissn

0006-2960

issn

1520-4995

journal_volume

27

pub_type

杂志文章

相关文献

BIOCHEMISTRY文献大全
  • Tritium isotope effects in adenosylcobalamin-dependent methylmalonyl-CoA mutase.

    abstract：:Methylmalonyl-CoA mutase from Propionibacterium shermanii is an adenosylcobalamin-dependent enzyme which catalyzes the reversible isomerization of methylmalonyl-CoA and succinyl-CoA. The rate of tritium loss from 5'-[3H]adenosylcobalamin during the enzymic reaction and the relative rates of tritium appearance in subst...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi961250o

    authors： Meier TW,Thomä NH,Leadlay PF

    更新日期：1996-09-10 00:00:00

  • Endogenous antithrombin associated with microvascular endothelium. Quantitative analysis in perfused rat hearts.

    abstract：:A recirculating Langendorff heart preparation is used to characterize the endogenous antithrombin associated reversibly with murine vascular endothelium. Rat hearts are perfused clear of blood and then recirculated with a physiological salt solution. Addition of heparin educes antithrombin activity continuously into t...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00169a024

    authors： Felsch JS,Owen WG

    更新日期：1994-01-25 00:00:00

  • Novel insights into the INK4-CDK4/6-Rb pathway: counter action of gankyrin against INK4 proteins regulates the CDK4-mediated phosphorylation of Rb.

    abstract：:The newly discovered oncogenic protein gankyrin, which contains six ankyrin repeats, has been reported to be involved in the phosphorylation and degradation of the retinoblastoma gene product, Rb. Using in vitro systems, we have identified a peptide fragment of gankyrin, 176LHLACDEERN185, which is responsible for bind...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi011550s

    authors： Li J,Tsai MD

    更新日期：2002-03-26 00:00:00

  • DNA strand arrangement within the SfiI-DNA complex: atomic force microscopy analysis.

    abstract：:The SfiI restriction enzyme binds to DNA as a tetramer holding two usually distant DNA recognition sites together before cleavage of the four DNA strands. To elucidate structural properties of the SfiI-DNA complex, atomic force microscopy (AFM) imaging of the complexes under noncleaving conditions (Ca2+ instead of Mg2...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi051767c

    authors： Lushnikov AY,Potaman VN,Oussatcheva EA,Sinden RR,Lyubchenko YL

    更新日期：2006-01-10 00:00:00

  • O(2)-evolving chlorite dismutase as a tool for studying O(2)-utilizing enzymes.

    abstract：:The direct interrogation of fleeting intermediates by rapid-mixing kinetic methods has significantly advanced our understanding of enzymes that utilize dioxygen. The gas's modest aqueous solubility (<2 mM at 1 atm) presents a technical challenge to this approach, because it limits the rate of formation and extent of a...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi201906x

    authors： Dassama LM,Yosca TH,Conner DA,Lee MH,Blanc B,Streit BR,Green MT,DuBois JL,Krebs C,Bollinger JM Jr

    更新日期：2012-02-28 00:00:00

  • Exchange mechanisms for hydrogen bonding protons of cytidylic and guanylic acids.

    abstract：:The pH dependence of buffer catalysis of exchange of the C-4 amino protons of cyclic cytosine 2',3'-monophosphate (cCMP) and the N-1 proton of cyclic guanosine 2',3'-monophosphate (cGMP) conforms to an exchange mechanism, in which protonation of the nucleobases at C(N-3) AND G(N-7) establishes the important intermedia...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00608a035

    authors： McConnell B

    更新日期：1978-07-25 00:00:00

  • Calcium-sensitive binding of heavy meromyosin to regulated actin requires light chain 2 and the head-tail junction.

    abstract：:Sedimentation in a preparative ultracentrifuge was used to determine the affinity of heavy meromyosin, HMM, for regulated actin, F-actin plus troponin-tropomyosin, in the presence of MgATP at pH 7.0, 20 degrees C, and mu = 18 mM. HMM was prepared from vertebrate striated muscle myosin by a mild chymotryptic digestion....

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00275a003

    authors： Wagner PD,Stone DB

    更新日期：1983-03-15 00:00:00

  • Time-resolved fluorescence resonance energy transfer shows that the bacterial multidrug ABC half-transporter BmrA functions as a homodimer.

    abstract：:Members of the ATP-binding cassette (ABC) transporters share the same basic architecture, with a four-core domain made of two transmembrane plus two nucleotide-binding domains. However, a supramolecular organization has been detected in some ABC transporters, which might be relevant to physiological regulation of subs...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi0482809

    authors： Dalmas O,Do Cao MA,Lugo MR,Sharom FJ,Di Pietro A,Jault JM

    更新日期：2005-03-22 00:00:00

  • Cobalt-cytochrome c. I. Preparation, properties, and enzymic activity.

    abstract：:An improved procedure for the preparation of cobalt-cytochrome c has been developed. Various factors influencing the cobalt insertion process are discussed. The optical spectra of cobalt-cytochrome c suggest a six-coordinated species. The spectral shifts occurring with oxidation-reduction are compared with those obser...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00687a003

    authors： Dickinson LC,Chien JC

    更新日期：1975-08-12 00:00:00

  • Effects of the DNA intercalators 4'-(9-acridinylamino)methanesulfon-m-anisidide and 2-methyl-9-hydroxyellipticinium on topoisomerase II mediated DNA strand cleavage and strand passage.

    abstract：:DNA topoisomerase II is believed to be the enzyme that produces the protein-associated DNA strand breaks observed in mammalian cell nuclei treated with various intercalating agents. Two intercalators--4'-(9-acridinylamino)methanesulfon-m-anisidide (m-AMSA, amsacrine) and 2-methyl-9-hydroxyellipticinium (2-Me-9-OH-E+)-...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00344a015

    authors： Pommier Y,Minford JK,Schwartz RE,Zwelling LA,Kohn KW

    更新日期：1985-11-05 00:00:00

  • Structural composition of canine secretory component and immunoglobulin A.

    abstract：:Dog serum and colostral immunoglobulin A (IgA) and free secretory component from colostrum were isolated using affinity chromatography. Both serum and colostral IgA showed similar susceptibility to reduction with dithiothreitol, but only colostral IgA released the additional subunit, bound secretory component. This re...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00684a010

    authors： Thompson RE,Reynolds HY,Waxdal MJ

    更新日期：1975-07-01 00:00:00

  • Direct identification of the agonist binding site in the human brain cholecystokininB receptor.

    abstract：:In investigating the agonist binding site of the human brain cholecystokininB receptor (CCKBR), we employed the direct protein chemical approach using a photoreactive tritiated analogue of sulfated cholecystokinin octapeptide, which contains the p-benzoylbenzoyl moiety at the N-terminus, followed by purification of th...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi990269z

    authors： Anders J,Blüggel M,Meyer HE,Kühne R,ter Laak AM,Kojro E,Fahrenholz F

    更新日期：1999-05-11 00:00:00

  • NMR structural studies of a 15-mer DNA sequence from a ras protooncogene, modified at the first base of codon 61 with the carcinogen 4-aminobiphenyl.

    abstract：:Proton NMR studies were conducted on the complementary 15-mer duplex d(5'-TACTCTTCTTGACCT).(5'-AGGTCAAGAAGAGTA) (designated as unmodified 15-mer duplex) spanning a portion of the mouse c-Ha-ras protooncogene centered around codon 61. Identical studies were carried out on the same sequence, after specific modification ...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00155a011

    authors： Cho BP,Beland FA,Marques MM

    更新日期：1992-10-13 00:00:00

  • Peroxynitrite induces long-lived tyrosyl radical(s) in oxyhemoglobin of red blood cells through a reaction involving CO2 and a ferryl species.

    abstract：:Peroxynitrite-mediated oxidative chemistry is currently the subject of intense investigation owing to the toxic side effects associated with nitric oxide overproduction. Using direct electron spin resonance spectroscopy (ESR) at 37 degrees C, we observed that in human erythrocytes peroxynitrite induced a long-lived si...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi982311g

    authors： Minetti M,Scorza G,Pietraforte D

    更新日期：1999-02-16 00:00:00

  • Phospholipid spin probes measure the effects of ethanol on the molecular order of liver microsomes.

    abstract：:Ethanol, in vitro, is known to perturb the molecular order of the phospholipids in biological membranes, while chronic ethanol exposure, in vivo, leads to resistance to disordering. Such changes have usually been measured by electron spin resonance, utilizing fatty acid spin probes. The use of such probes is controver...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00346a012

    authors： Taraschi TF,Wu A,Rubin E

    更新日期：1985-12-03 00:00:00

  • Spectroscopic Determination of Distinct Heme Ligands in Outer-Membrane Receptors PhuR and HasR of Pseudomonas aeruginosa.

    abstract：:Pseudomonas aeruginosa PAO1 encodes two outer membrane receptors, PhuR (Pseudomonas heme uptake) and HasR (heme assimilation system). The HasR receptor acquires heme through interaction with a secreted hemophore, HasAp. The non-hemophore-dependent PhuR is encoded along with proteins required for heme translocation int...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/acs.biochem.5b00017

    authors： Smith AD,Modi AR,Sun S,Dawson JH,Wilks A

    更新日期：2015-04-28 00:00:00

  • Biochemical Characterization of Yeast Xrn1.

    abstract：:Messenger RNA degradation is an important component of overall gene expression. During the final step of eukaryotic mRNA degradation, exoribonuclease 1 (Xrn1) carries out 5' → 3' processive, hydrolytic degradation of RNA molecules using divalent metal ion catalysis. To initiate studies of the 5' → 3' RNA decay machine...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/acs.biochem.9b01035

    authors： Langeberg CJ,Welch WRW,McGuire JV,Ashby A,Jackson AD,Chapman EG

    更新日期：2020-04-21 00:00:00

  • Leaving group dependence and proton inventory studies of the phosphorylation of a cytoplasmic phosphotyrosyl protein phosphatase from bovine heart.

    abstract：:The kcat and Km values for the bovine heart low molecular weight phosphotyrosyl protein phosphatase catalyzed hydrolysis of 16 aryl phosphate monoesters and of five alkyl phosphate monoesters having the structure Ar(CH2)nOPO3H2 (n = 1-5) were measured at pH 5.0 and 37 degrees C. With the exception of alpha-naphthyl ph...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00101a006

    authors： Zhang ZY,Van Etten RL

    更新日期：1991-09-17 00:00:00

  • pH dependence and structural interpretation of the reactions of Coprinus cinereus peroxidase with hydrogen peroxide, ferulic acid, and 2,2'-azinobis.

    abstract：:Steady-state and transient-state analysis of Coprinus cinereus peroxidase, CIP (identical to Arthromyces ramosus peroxidase), was used to characterize the kinetics of the three fundamental steps in heme peroxidase catalysis: compound I (cpd I) formation, cpd I reduction, and compound II (cpd II) reduction. The rate co...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi970387r

    authors： Abelskov AK,Smith AT,Rasmussen CB,Dunford HB,Welinder KG

    更新日期：1997-08-05 00:00:00

  • Metal ion binding to dog osteocalcin studied by 1H NMR spectroscopy.

    abstract：:One-dimensional 1H NMR was employed to study the effects of Ca2+ and Lu3+ binding on the apo and calcium-saturated forms of dog bone Gla protein (BGP, osteocalcin). Titration of apo dog BGP with Ca2+ in 20 mM NaCl showed spectral perturbations consistent with the binding of 5 mol equiv of calcium in the NMR slow-excha...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00093a012

    authors： Isbell DT,Du S,Schroering AG,Colombo G,Shelling JG

    更新日期：1993-10-26 00:00:00

  • Phospholipid asymmetry in large unilamellar vesicles induced by transmembrane pH gradients.

    abstract：:The influence of membrane pH gradients on the transbilayer distribution of some common phospholipids has been investigated. We demonstrate that the transbilayer equilibrium of the acidic phospholipids egg phosphatidylglycerol (EPG) and egg phosphatidic acid (EPA) can be manipulated by membrane proton gradients, wherea...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00436a009

    authors： Hope MJ,Redelmeier TE,Wong KF,Rodrigueza W,Cullis PR

    更新日期：1989-05-16 00:00:00

  • Structural alterations of Lamin A protein in dilated cardiomyopathy.

    abstract：:Lamin A protein, encoded by the LMNA gene, belongs to the type V intermediate filament protein family and is a major nuclear protein component of higher metazoan organisms, including humans. Lamin A along with B-type lamins impart structural rigidity to the nucleus by forming a lamina that is closely apposed to the in...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi400337t

    authors： Bhattacharjee P,Banerjee A,Banerjee A,Dasgupta D,Sengupta K

    更新日期：2013-06-18 00:00:00

  • Topography of the Dictyostelium discoideum plasma membrane: analysis of membrane asymmetry and intermolecular disulfide bonds.

    abstract：:Through the application of a unique method for isolating plasma membranes, it was possible to specifically iodinate cytoplasm-exposed plasma membrane proteins in vegetative cells of the cellular slime mold Dictyostelium discoideum. The original procedure [Chaney, L. K., & Jacobson, B. S. (1983) J. Biol. Chem. 258, 100...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00389a041

    authors： Shiozawa JA,Jelenska MM,Jacobson BS

    更新日期：1987-07-28 00:00:00

  • Peptidylglycine alpha-hydroxylating monooxygenase: active site residues, disulfide linkages, and a two-domain model of the catalytic core.

    abstract：:Peptidylglycine alpha-hydroxylating monooxygenase (PHM) is a copper, ascorbate, and molecular oxygen dependent enzyme that catalyzes the first step leading to the C-terminal amidation of glycine-extended peptides. The catalytic core of PHM (PHMcc), refined to residues 42-356 of the PHM protein, was expressed at high l...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi9708747

    authors： Kolhekar AS,Keutmann HT,Mains RE,Quon AS,Eipper BA

    更新日期：1997-09-09 00:00:00

  • Calmodulin-dependent protein phosphatase: a developmental study.

    abstract：:Calmodulin-dependent protein phosphatase, one of the major calmodulin-binding proteins in bovine brain, dephosphorylates casein with a specific activity of 15 nmol mg-1 min-1 at 30 degrees C. The stimulation of phosphatase activity by calmodulin is reversed by ethylene glycol bis (beta-aminoethyl ether)-N,N,N',N'-tetr...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00284a014

    authors： Tallant EA,Cheung WY

    更新日期：1983-07-19 00:00:00

  • Studies on transcription of 3'-extended templates by mammalian RNA polymerase II. Parameters that affect the initiation and elongation reactions.

    abstract：:Addition of short sequences of dCMP residues to the 3'-OH end of duplex linear DNAs allows rapid and efficient transcription to be initiated at these sites by purified mammalian RNA polymerase II [Kadesch, T. R., & Chamberlin, M. J. (1982) J. Biol. Chem. 257, 5286-5295]. The use of such tailed DNA templates should all...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00330a019

    authors： Dedrick RL,Chamberlin MJ

    更新日期：1985-04-23 00:00:00

  • Is the PTPase-vanadate complex a true transition state analogue?

    abstract：:Vanadate can often bind to phosphoryl transfer enzymes to form a trigonal-bipyramidal structure at the active site. The enzyme-vanadate dissociation constants in these enzymes are much lower than those for phosphate. Therefore, enzyme-bound vanadate moieties are often considered as transition state analogues. To test ...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi016097z

    authors： Deng H,Callender R,Huang Z,Zhang ZY

    更新日期：2002-05-07 00:00:00

  • Selective inhibition by methoxyamine of the apurinic/apyrimidinic endonuclease activity associated with pyrimidine dimer-DNA glycosylases from Micrococcus luteus and bacteriophage T4.

    abstract：:The UV endonucleases [endodeoxyribonuclease (pyrimidine dimer), EC 3.1.25.1] from Micrococcus luteus and bacteriophage T4 possess two catalytic activities specific for the site of cyclobutane pyrimidine dimers in UV-irradiated DNA: a DNA glycosylase that cleaves the 5'-glycosyl bond of the dimerized pyrimidines and an...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi00386a011

    authors： Liuzzi M,Weinfeld M,Paterson MC

    更新日期：1987-06-16 00:00:00

  • Decrypting the structural, dynamic, and energetic basis of a monomeric kinesin interacting with a tubulin dimer in three ATPase states by all-atom molecular dynamics simulation.

    abstract：:We have employed molecular dynamics (MD) simulation to investigate, with atomic details, the structural dynamics and energetics of three major ATPase states (ADP, APO, and ATP state) of a human kinesin-1 monomer in complex with a tubulin dimer. Starting from a recently solved crystal structure of ATP-like kinesin-tubu...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi501056h

    authors： Chakraborty S,Zheng W

    更新日期：2015-01-27 00:00:00

  • Vinca alkaloid-induced tubulin spiral formation correlates with cytotoxicity in the leukemic L1210 cell line.

    abstract：:The ability of a class of C-20' modified vinca alkaloid congeners to induce tubulin spiral formation was investigated relative to their ability to inhibit microtubule assembly, their cytotoxicity against a leukemic cell line, L1210, and their measured and calculated partition coefficients. These studies were prompted ...

    journal_title：Biochemistry

    pub_type： 杂志文章

    doi：10.1021/bi001038r

    authors： Lobert S,Fahy J,Hill BT,Duflos A,Etievant C,Correia JJ

    更新日期：2000-10-03 00:00:00