NMR study of the inhibition of pepsin by glyoxal inhibitors: mechanism of tetrahedral intermediate stabilization by the aspartyl proteases.

Abstract:

:Z-Ala-Ala-Phe-glyoxal (where Z is benzyloxycarbonyl) has been shown to be a competitive inhibitor of pepsin with a Ki = 89 +/- 24 nM at pH 2.0 and 25 degrees C. Both the ketone carbon (R13COCHO) and the aldehyde carbon (RCO13CHO) of the glyoxal group of Z-Ala-Ala-Phe-glyoxal have been 13C-enriched. Using 13C NMR, it has been shown that when the inhibitor is bound to pepsin, the glyoxal keto and aldehyde carbons give signals at 98.8 and 90.9 ppm, respectively. This demonstrates that pepsin binds and preferentially stabilizes the fully hydrated form of the glyoxal inhibitor Z-Ala-Ala-Phe-glyoxal. From 13C NMR pH studies with glyoxal inhibitor, we obtain no evidence for its hemiketal or hemiacetal hydroxyl groups ionizing to give oxyanions. We conclude that if an oxyanion is formed its pKa must be >8.0. Using 1H NMR, we observe four hydrogen bonds in free pepsin and in pepsin/Z-Ala-Ala-Phe-glyoxal complexes. In the pepsin/pepstatin complex an additional hydrogen bond is formed. We examine the effect of pH on hydrogen bond formation, but we do not find any evidence for low-barrier hydrogen bond formation in the inhibitor complexes. We conclude that the primary role of hydrogen bonding to catalytic tetrahedral intermediates in the aspartyl proteases is to correctly orientate the tetrahedral intermediate for catalysis.

journal_name

Biochemistry

journal_title

Biochemistry

authors

Cosgrove S,Rogers L,Hewage CM,Malthouse JP

doi

10.1021/bi701000k

subject

Has Abstract

pub_date

2007-10-02 00:00:00

pages

11205-15

issue

39

eissn

0006-2960

issn

1520-4995

journal_volume

46

pub_type

杂志文章
  • Influence of tertiary structure domain properties on the functionality of apolipoprotein A-I.

    abstract::The tertiary structure of apolipoprotein (apo) A-I and the contributions of structural domains to the properties of the protein molecule are not well defined. We used a series of engineered human and mouse apoA-I molecules in a range of physical-biochemical measurements to address this issue. Circular dichroism measur...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi702332b

    authors: Tanaka M,Koyama M,Dhanasekaran P,Nguyen D,Nickel M,Lund-Katz S,Saito H,Phillips MC

    更新日期:2008-02-19 00:00:00

  • Interactions between the mitochondrial adenosinetriphosphatase and periodate-oxidized adenosine 5'-triphosphate, an affinity label for adenosine 5'-triphosphate binding sites.

    abstract::Periodate-oxidized ATP (o-ATP) was prepared as an affinity label of nucleotide binding sites on the chloroform-released ox heart mitochondrial ATPase. In the presence of MgSO4, o-ATP is a substrate for the ATPase. It can act as a reversible, competitive inhibitor of ATPase activity and can also induce an irreversible ...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00260a025

    authors: Lowe PN,Beechey RB

    更新日期:1982-08-17 00:00:00

  • Effects of biological oxidants on the catalytic activity and structure of group VIA phospholipase A2.

    abstract::Group VIA phospholipase A(2) (iPLA(2)beta) is expressed in phagocytes, vascular cells, pancreatic islet beta-cells, neurons, and other cells and plays roles in transcriptional regulation, cell proliferation, apoptosis, secretion, and other events. A bromoenol lactone (BEL) suicide substrate used to study iPLA(2)beta f...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi060502a

    authors: Song H,Bao S,Ramanadham S,Turk J

    更新日期:2006-05-23 00:00:00

  • Role of isomerization of initial complexes in the binding of inhibitors to dihydrofolate reductase.

    abstract::Stopped-flow measurements of protein fluorescence quenching when methotrexate (MTX) binds to dihydrofolate reductase (isoenzyme II) of Streptococcus faecium (SFDHFR II) analyze as the sum of two differentials: a rapid binding phase and a second phase for which the observed rate constant is independent of methotrexate ...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00339a010

    authors: Blakley RL,Cocco L

    更新日期:1985-08-27 00:00:00

  • Mechanistic insights from the binding of substrate and carbocation intermediate analogues to aristolochene synthase.

    abstract::Aristolochene synthase, a metal-dependent sesquiterpene cyclase from Aspergillus terreus, catalyzes the ionization-dependent cyclization of farnesyl diphosphate (FPP) to form the bicyclic eremophilane (+)-aristolochene with perfect structural and stereochemical precision. Here, we report the X-ray crystal structure of...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi400691v

    authors: Chen M,Al-lami N,Janvier M,D'Antonio EL,Faraldos JA,Cane DE,Allemann RK,Christianson DW

    更新日期:2013-08-13 00:00:00

  • O-phosphoserine content of intermediate filament subunits.

    abstract::Purified subunits of intermediate filaments obtained from a variety of tissues and cell types contain O-phosphoserine and, in some cases, smaller amounts of O-phosphothreonine. The O-phosphoserine content was estimated by reaction of performic acid oxidized subunits with methylamine in NaOH. Decamin of BHK-21 and CHO ...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00530a030

    authors: Steinert PM,Wantz ML,Idler WW

    更新日期:1982-01-05 00:00:00

  • Multiple equilibria binding treatment of lipid and detergent interactions with membrane proteins. Application to cytochrome c oxidase solubilized in cholate.

    abstract::A modified multiple binding equilibria treatment is presented that allows determination of thermodynamic parameters of the interaction of phospholipids with integral membrane proteins solubilized in excess detergent. Lipid binding is modeled as a series of exchange reactions between lipid molecules and detergent molec...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00376a019

    authors: Volwerk JJ,Mrsny RJ,Patapoff TW,Jost PC,Griffith OH

    更新日期:1987-01-27 00:00:00

  • Interaction between SecA and SecYEG in micellar solution and formation of the membrane-inserted state.

    abstract::Preprotein translocation in Escherichia coli is mediated by the translocase with SecA as peripheral ATPase and SecY, SecE, and SecG as membrane domain. To facilitate large-scale purification of the SecYEG heterotrimer, SecY was fused at its amino terminus with a hexahistidine tag and co-overexpressed with SecE and Sec...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi972105t

    authors: van der Does C,Manting EH,Kaufmann A,Lutz M,Driessen AJ

    更新日期:1998-01-06 00:00:00

  • Kinetics and mechanism of transitions involving the lamellar, cubic, inverted hexagonal, and fluid isotropic phases of hydrated monoacylglycerides monitored by time-resolved X-ray diffraction.

    abstract::A study of the dynamics and mechanism of the various thermotropic phase transitions undergone by the hydrated monoacylglycerides monoolein and monoelaidin, in the temperature range of 20-120 degrees C and from 0 to 5 M NaCl, has been undertaken. Measurements were made by using time-resolved X-ray diffraction at the Co...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00394a008

    authors: Caffrey M

    更新日期:1987-10-06 00:00:00

  • Potassium flux through gramicidin ion channels is augmented in vesicles comprised of plasmenylcholine: correlations between gramicidin conformation and function in chemically distinct host bilayer matrices.

    abstract::The functional role of distinct phospholipid subclasses and molecular species in modulating gramicidin-mediated K+ flux was characterized through quantification of changes in the fluorescence intensity of ion specific fluorescent probes encapsulated inside vesicles comprised of individual molecular species of plasmeny...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00022a008

    authors: Chen X,Gross RW

    更新日期:1995-06-06 00:00:00

  • Complexity of Bovine Rhodopsin Activation Revealed at Low Temperature and Alkaline pH.

    abstract::The late intermediates involved in the activation mechanism of bovine rhodopsin are investigated by time-resolved optical absorption spectroscopy. Measurements from 10 μs to 200 ms after photolysis were carried out on membrane suspensions of bovine rhodopsin at a temperature of 15 °C and at pH of 7.3, 8.0, and 8.7. Th...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/acs.biochem.6b00687

    authors: Szundi I,Funatogawa C,Kliger DS

    更新日期:2016-09-13 00:00:00

  • Extended X-ray absorption fine structure of copper in CuA-depleted, p-(hydroxymercuri)benzoate-modified, and native cytochrome c oxidase.

    abstract::Cytochrome c oxidase contains four redox-active metal centers: two heme irons, cytochromes a and a3, and two copper ions, CuA and CuB. Due to the paucity of spectroscopic signatures for both copper sites in cytochrome c oxidase, the ligands and structures for these sites have remained ambiguous. The specific depletion...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00382a005

    authors: Li PM,Gelles J,Chan SI,Sullivan RJ,Scott RA

    更新日期:1987-04-21 00:00:00

  • Structural coupling between the oxygen-evolving Mn cluster and a tyrosine residue in photosystem II as revealed by Fourier transform infrared spectroscopy.

    abstract::The flash-induced Fourier transform infrared (FTIR) difference spectrum of the oxygen-evolving Mn cluster upon S1-to-S2 transition (S2/S1 spectrum) was measured using photosystem II (PS II) core complexes of Synechocystis 6803 in which tyrosine residues were specifically labeled with 13C at the ring-4 position. The do...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi971760y

    authors: Noguchi T,Inoue Y,Tang XS

    更新日期:1997-12-02 00:00:00

  • Energetic effects of magnesium in the recognition of adenosine nucleotides by the F(1)-ATPase beta subunit.

    abstract::Nucleotide-induced conformational changes of the catalytic beta subunits play a crucial role in the rotary mechanism of F(1)-ATPase. To gain insights into the energetic bases that govern the recognition of nucleotides by the isolated beta subunit from thermophilic Bacillus PS3 (Tbeta), the binding of this monomer to M...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi1006767

    authors: Pulido NO,Salcedo G,Pérez-Hernández G,José-Núñez C,Velázquez-Campoy A,García-Hernández E

    更新日期:2010-06-29 00:00:00

  • Generation and Comparative Kinetic Analysis of New Glycosynthase Mutants from Streptococcus pyogenes Endoglycosidases for Antibody Glycoengineering.

    abstract::Chemoenzymatic glycan remodeling by endoglycosidase-catalyzed deglycosylation and reglycosylation is emerging as an attractive approach for producing homogeneous glycoforms of antibodies, and the success of this approach depends on the discovery of efficient endoglycosidases and their glycosynthase mutants. We report ...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/acs.biochem.8b00719

    authors: Tong X,Li T,Li C,Wang LX

    更新日期:2018-09-04 00:00:00

  • A coupled dinuclear iron cluster that is perturbed by substrate binding in myo-inositol oxygenase.

    abstract::myo-Inositol oxygenase (MIOX) uses iron as its cofactor and dioxygen as its cosubstrate to effect the unique, ring-cleaving, four-electron oxidation of its cyclohexan-(1,2,3,4,5,6-hexa)-ol substrate to d-glucuronate. The nature of the iron cofactor and its interaction with the substrate, myo-inositol (MI), have been p...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi0519607

    authors: Xing G,Hoffart LM,Diao Y,Prabhu KS,Arner RJ,Reddy CC,Krebs C,Bollinger JM Jr

    更新日期:2006-05-02 00:00:00

  • Evidence for a slow tertiary relaxation in the reaction of tert-butyl isocyanide with horseradish peroxidase.

    abstract::The kinetics of tert-butyl isocyanide binding to the heme protein horseradish peroxidase (HRP) at 22 degrees C was examined on all time scales, from minutes to picoseconds, in aqueous borate buffer at pH 9.08. Unlike myoglobin (Mb) or hemoglobin, HRP shows two bimolecular ligand binding processes. For comparison, bind...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi9518149

    authors: Bandyopadhyay D,Walda KN,Grogan TM,Magde D,Traylor TG,Sharma VS

    更新日期:1996-02-06 00:00:00

  • Probing receptor binding activity of interleukin-8 dimer using a disulfide trap.

    abstract::Interleukin-8 (IL-8), a member of the chemokine superfamily, exists as both monomers and dimers, and mediates its function by binding to neutrophil CXCR1 and CXCR2 receptors that belong to the G protein-coupled receptor class. It is now well established that the monomer functions as a high-affinity ligand, but the bin...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi0605944

    authors: Rajarathnam K,Prado GN,Fernando H,Clark-Lewis I,Navarro J

    更新日期:2006-06-27 00:00:00

  • Ciliary dynein conformational changes as evidenced by the extrinsic fluorescent probe 8-anilino-1-naphthalenesulfonate.

    abstract::The binding of 8-anilino-1-naphthalenesulfonate (ANS) to ciliary dynein ATPase leads to a marked increase in the dye's fluorescence intensity, accompanied by a blue shift in the observed fluorescence emission maximum. We found that dynein has 37 +/- 3 ANS binding sites and that experimentally applied ANS concentration...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00347a012

    authors: Saucier AC,Mariotti S,Anderson SA,Purich DL

    更新日期:1985-12-17 00:00:00

  • Mechanism of the quorum-quenching lactonase (AiiA) from Bacillus thuringiensis. 2. Substrate modeling and active site mutations.

    abstract::The N-acyl- l-homoserine lactone hydrolases (AHL lactonases) have attracted considerable attention because of their ability to quench AHL-mediated quorum-sensing pathways in Gram-negative bacteria and because of their relation to other enzymes in the metallo-beta-lactamase superfamily. To elucidate the detailed cataly...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi8003704

    authors: Momb J,Wang C,Liu D,Thomas PW,Petsko GA,Guo H,Ringe D,Fast W

    更新日期:2008-07-22 00:00:00

  • Kinetics of binding of chicken cystatin to papain.

    abstract::The kinetics of binding of chicken cystatin to papain were studied by stopped-flow fluorometry under pseudo-first-order conditions, i.e., with an excess of inhibitor. All reactions showed first-order behavior, and the observed pseudo-first-order rate constant increased linearly with the cystatin concentration up to th...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00430a022

    authors: Björk I,Alriksson E,Ylinenjärvi K

    更新日期:1989-02-21 00:00:00

  • Linkage between operator binding and dimer to octamer self-assembly of bacteriophage lambda cI repressor.

    abstract::Cooperative binding of the bacteriophage lambda cI repressor dimer to specific sites of the phage operators OR and OL controls the developmental state of the phage. Cooperativity has long been thought to be mediated by self-assembly of repressor dimers to form tetramers which can bind simultaneously to adjacent operat...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi971368k

    authors: Rusinova E,Ross JB,Laue TM,Sowers LC,Senear DF

    更新日期:1997-10-21 00:00:00

  • MAP4 is the in vivo substrate for CDC2 kinase in HeLa cells: identification of an M-phase specific and a cell cycle-independent phosphorylation site in MAP4.

    abstract::We reported previously that cdc2 kinase decreased the microtubule-stabilizing ability of a major HeLa cell microtubule-associated protein, MAP4, by phosphorylation in vitro [Ookata, K., et al. (1995) J. Cell Biol. 128, 849-862]. An important question raised by this study is whether MAP4 is indeed phosphorylated by cdc...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi971251w

    authors: Ookata K,Hisanaga S,Sugita M,Okuyama A,Murofushi H,Kitazawa H,Chari S,Bulinski JC,Kishimoto T

    更新日期:1997-12-16 00:00:00

  • Structural symmetry: the three-dimensional structure of Haemophilus influenzae diaminopimelate epimerase.

    abstract::The Haemophilus influenzae diaminopimelate epimerase was cloned, expressed, purified, and crystallized in the C2221 space group (a = 102.1 A, b = 115.4 A, c = 66.3 A, alpha = beta = gamma = 90 degrees). The three-dimensional structure was solved to 2.7 A using a single Pt derivative and the Se-Met-substituted enzyme t...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi982138o

    authors: Cirilli M,Zheng R,Scapin G,Blanchard JS

    更新日期:1998-11-24 00:00:00

  • Independent functions for the N- and C-termini in the overlap region of tropomyosin.

    abstract::Tropomyosin (TM) is a coiled-coil that binds head-to-tail along the helical actin filament. The ends of 284-residue tropomyosins are believed to overlap by about nine amino acids. The present study investigates the function of the N- and C-terminal overlap regions. Recombinant tropomyosins were produced in Escherichia...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi000242b

    authors: Moraczewska J,Hitchcock-DeGregori SE

    更新日期:2000-06-13 00:00:00

  • Intermediates in excision repair by human cells: use of S1 nuclease and benzoylated naphthoylated cellulose to reveal single-strand breaks.

    abstract::Treatment of human lymphoid cells with methyl methanesulfonate (MMS) results in discontinuities in cellular DNA. Discontinuities can be detected by sedimentation through neutral sucrose gradients after digestion with S1 nuclease on top of the gradient and by benzoylated naphthoylated diethylaminoethylcellulose (BND-ce...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00639a024

    authors: Karran P,Higgins NP,Strauss B

    更新日期:1977-10-04 00:00:00

  • Synthesis and study of phosphoenolthiopyruvate.

    abstract::Phosphoenolthiopyruvate, the analogue of phosphoenolpyruvate in which the bridging oxygen of the phosphate ester is replaced by sulfur, has been synthesized from methyl acrylate and dimethyl (chlorothio)phosphonate. The compound is a substrate for alkaline phosphatase, pyruvate kinase, enolase, and phosphoenolpyruvate...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00404a038

    authors: Sikkema KD,O'Leary MH

    更新日期:1988-02-23 00:00:00

  • Infrared spectroscopic study of the secondary structure of melittin in water, 2-chloroethanol, and phospholipid bilayer dispersions.

    abstract::The conformations of melittin, an amphipathic polypeptide consisting of 26 amino acid residues, and its hydrophobic (residues 1--19) and hydrophilic (residues 20--26) fragments were examined in various solvent systems, including H2O, 2H2O, 2-chloroethanol, and 1,2-dimyristoylphosphatidylcholine (DMPC) multilayers, by ...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00539a006

    authors: Lavialle F,Adams RG,Levin IW

    更新日期:1982-05-11 00:00:00

  • Implication of His68 in the substrate site of Bacillus subtilis adenylosuccinate lyase by mutagenesis and affinity labeling with 2-[(4-bromo-2,3-dioxobutyl)thio]adenosine 5'-monophosphate.

    abstract::Adenylosuccinate lyase of Bacillus subtilis is inactivated by 2-[(4-bromo-2,3-dioxobutyl)thio]adenosine 5'-monophosphate (2-BDB-TAMP) at pH 7.0. As the reagent concentration is increased, a maximum rate constant is approached, indicative of reversible enzyme-reagent complex formation (KR = 68 +/- 9 microM) prior to ir...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi9805339

    authors: Lee TT,Worby C,Bao ZQ,Dixon JE,Colman RF

    更新日期:1998-06-09 00:00:00

  • Biochemical and kinetic characteristics of the interaction of the antitumor antibiotic sparsomycin with prokaryotic and eukaryotic ribosomes.

    abstract::Using 125I-labeled phenol-alanine sparsomycin, an analogue of sparsomycin having higher biological activity than the unmodified antibiotic, we studied the requirements and the characteristics of its interaction with the ribosome. The drug does not bind to either isolated ribosomal subunits or reconstituted whole ribos...

    journal_title:Biochemistry

    pub_type: 杂志文章

    doi:10.1021/bi00104a011

    authors: Lazaro E,van den Broek LA,San Felix A,Ottenheijm HC,Ballesta JP

    更新日期:1991-10-08 00:00:00