Inactivation of class I fructose diphosphate aldolases by the substrate analog N-bromoacetylethanolamine phosphate.

Abstract N-Bromoacetylethanolamine phosphate, prepared by the bromoacetylation of ethanolamine phosphate, has been tested as an active site-specific reagent for rabbit and rat muscle fructose diphosphate aldolases. The reagent inactivates both enzymes, and inactivation is prevented by substrates or competitive inhibitors. Loss of activity is pseudo-first order until the later stages of inactivation, and a rate-saturation effect is observed as the reagent concentration is increased. The stoichiometry of the reaction has been determined with 14C-labeled reagent. Inactivated enzyme contains 2 to 2.5 molar eq of reagent per mole of aldolase subunit, whereas the enzyme modified in the presence of a competitive inhibitor contains only 1 to 1.5 eq of reagent. Thus, alterations in the catalytic properties of aldolase that accompany its modification apparently reflect the alkylation of a single, essential residue. Characterization of the protein derivatives reveals that the preservation of enzymatic activity is due primarily to the protection of a histidyl residue. This observation supports earlier evidence for the involvement of a histidyl residue in aldolase activity.

• Publication year

  1973

• Venue

  Journal of Biological Chemistry

• Publication date

  1973-12-10

• Fields of study

  Medicine, Chemistry

• Identifiers
  DOI 10.1016/s0021-9258(19)43219-5PMID 4752953
• External record

  Open on Semantic Scholar

• Source metadata

  Semantic Scholar, PubMed

• No claims are published for this paper.

• No concepts are published for this paper.

• Haloacetol phosphates. Identification of the sulfhydryl group of rabbit muscle aldolase alkylated by chloroacetol phosphate.

  1973cited by this paper
• Structural comparisons between the class I fructose diphosphate aldolases from Micrococcus aerogenes and rabbit.

  1973cited by this paper
• Covalent attachment of a peptidyl-transfer RNA analog to the 50S subunit of Escherichia coli ribosomes.

  1972cited by this paper
• The number, location, and reactivity of the cysteine residues of sturgeon muscle aldolase.

  1972cited by this paper
• Haloacetol phosphates. Selective alkylation of sulfhydryl groups of rabbits muscle aldolase by chloroacetol phosphate.

  1972cited by this paper
• Fluorescence studies of the binding of alkyl and aryl phosphates to rat muscle aldolase.

  1971cited by this paper
• Identification of a cysteinyl residue involved in the activity of rabbit muscle aldolase.

  1970cited by this paper
• The reactivity of thiol groups and the subunit structure of aldolase.

  1970cited by this paper
• Haloacetol phosphates. Potential active-site reagents for aldolase, triose phosphate isomerase, and glycerophosphate dehydrogenase. I. Preparation and properties.

  1970cited by this paper
• Photosensitization of specific histidine residues of rabbit muscle and spinach leaf aldolases by pyridoxal phosphate.

  1970cited by this paper
• Participation of cysteinyl residues in the structure and function of muscle aldolase. Characterization of mixed disulfide derivatives.

  1970cited by this paper
• Studies on the interaction of aldolase with substrate analogues.

  1969cited by this paper
• A functional role of metal ions in a class II aldolase.

  1969cited by this paper
• High recovery of tryptophan from acid hydrolysates of proteins.

  1969cited by this paper
• A comparative study of the structure of muscle fructose 1,6-diphosphate aldolases.

  1969cited by this paper
• Ioslation of fructose diphosphate aldolases A, B, and C.

  1969cited by this paper
• Analysis of carboxymethylated residues in proteins by an isotopic method, and its application to the bromoacetate-ribonuclease reaction.

  1968cited by this paper
• Reaction of an aldolase-substrate intermediate with tetranitromethane.

  1968cited by this paper
• Labeling of the active site of aldolase with glyceraldehyde 3-phosphate and erythrose 4-phosphate.

  1968cited by this paper
• Catalysis of an oxygen-exchange reaction of fructose 1,6-diphosphate and fructose 1-phosphate with water by rabbit muscle aldolase.

  1968cited by this paper
• The subunit structure of mammalian fructose diphosphate aldolase.

  1967cited by this paper
• Bromopyruvate inactivation of 2-keto-3-deoxy-6-phosphogluconic aldolase. I. Kinetic evidence for active site specificity.

  1967cited by this paper
• The number of polypeptide chains in rabbit muscle aldolase.

  1966cited by this paper
• Specific anion binding to fructose diphosphate aldolase from rabbit muscle.

  1966cited by this paper
• The synthesis of N-acetylphenylalanyl-sRNA.

  1966cited by this paper
• An exploration of the active site of aldolase using structural analogs of fructose diphosphate.

  1965cited by this paper
• MECHANISM OF THE ALDOLASE REACTION.

  1965cited by this paper
• THE SPECTROPHOTOMETRIC TITRATION OF THE SULFHYDRYL AND PHENOLIC GROUPS OF ALDOLASE.

  1964cited by this paper
• EVOLUTION OF ALDOLASE.

  1964cited by this paper
• COMPARATIVE STUDIES OF LIVER AND MUSCLE ALDOLASE. II. IMMUNOCHEMICAL AND CHROMATOGRAPHIC DIFFERENTIATION.

  1963cited by this paper
• THE MECHANISM OF ACTION OF ALDOLASES. IV. LYSINE AS THE SUBSTRATE-BINDING SITE.

  1963cited by this paper
• Kinetic properties of native and carboxy-peptidase-altered rabbit muscle aldolase.

  1963cited by this paper
• The catalytic activity of carboxypeptidase-degraded aldolase.

  1959cited by this paper
• Tissue sulfhydryl groups.

  1959cited by this paper
• The reaction of iodoacetate with methionine.

  1959cited by this paper
• The mechanism of action of fructose diphosphate aldolase.

  1958cited by this paper
• Absolute Configuration of Enantiomorphic Carbanions involved in the Aldolase and Triose Phosphate Isomerase Reactions

  1958cited by this paper
• Studies on the mechanism on the aldolase reaction; isotope exchange reactions of muscle and yeast aldolase.

  1958cited by this paper
• On the cysteine content of human hemoglobin.

  1958cited by this paper
• The oxidation of ribonuclease with performic acid.

  1956cited by this paper

• The potential utility of PFKFB3 as a therapeutic target

  2018cites this paper
• Probing the location and function of the conserved histidine residue of phosphoglucose isomerase by using an active site directed inhibitor N‐bromoacetylethanolamine phosphate

  2008cites this paper
• The Crystal Structure of Phosphoglucose Isomerase/Autocrine Motility Factor/Neuroleukin Complexed with Its Carbohydrate Phosphate Inhibitors Suggests Its Substrate/Receptor Recognition*

  2000cites this paper
• Inhibition of Tumor Cell Growth by A Specific 6-Phosphofructo-2-kinase Inhibitor, N-Bromoacetylethanolamine Phosphate, and Its Analogues

  2000cites this paper
• N-Bromoacetylethanolamine phosphate as a probe for the identification of a liver microsomal glucose-6-phosphate transporter peptide in rats and Ehrlich ascites tumor-bearing mice.

  2000cites this paper
• Histone II-A activates the glucose-6-phosphatase system without microsomal membrane permeabilization.

  1998cites this paper
• Inhibition of fructose-6-phosphate,2-kinase by N-bromoacetylethanolamine phosphate in vitro and in vivo.

  1997cites this paper
• Inhibition of the glucose-6-phosphatase system by N-bromoacetylethanolamine phosphate, a potential affinity label for auxiliary proteins.

  1996cites this paper
• Hexose phosphate binding sites of fructose 6-phosphate,2-kinase:fructose 2,6-bisphosphatase.

  1995cites this paper
• Suppression of phase separation in solutions of bovine gamma IV-crystallin by polar modification of the sulfur-containing amino acids.

  1991cites this paper
• Affinity labeling of spinach phosphoribulokinase subsequent toS-methylation at Cys16

  1990cites this paper
• Catalytic site of rat liver and bovine heart fructose-6-phosphate,2-kinase:fructose-2,6-bisphosphatase. Identification of fructose 6-phosphate binding site.

  1989cites this paper
• The Thioredoxin Binding Site of Phosphoribulokinase Overlaps the Catalytic Site

  1987cites this paper
• Evidence for catalytic site cysteine and histidine by chemical modification of β-hydroxy-β-methylglutaryl-coenzyme a reductase

  1986cites this paper
• Evidence for catalytic site cysteine and histidine by chemical modification of beta-hydroxy-beta-methylglutaryl-coenzyme A reductase.

  1986cites this paper
• Evidence for a reactive cysteine at the nucleotide binding site of spinach ribulose-5-phosphate kinase.

  1985cites this paper
• Purification of the catalytically active phosphorylated form of insulin receptor kinase by affinity chromatography with O-phosphotyrosyl-binding antibodies.

  1985cites this paper
• Hexose phosphate binding sites of fructose-6-phosphate,2-kinase:fructose-2,6-bisphosphatase. Interaction with N-bromoacetylethanolamine phosphate and 3-bromo-1,4-dihydroxy-2-butanone 1,4-bisphosphate.

  1984cites this paper
• Chapter 8 Enzyme reactions involving imine formation

  1984cites this paper
• Suicide inactivation of fructose-1,6-bisphosphate aldolase.

  1984cites this paper
• 2-(4-Bromoacetamido)anilino-2-deoxypentitol 1,5-bisphosphate, a new affinity label for ribulose bisphosphate carboxylase/oxygenase from Rhodospirillum rubrum. Determination of reaction parameters and characterization of an active site peptide.

  1984cites this paper
• 2-Bromoacetylaminopentitol 1,5-bisphosphate as an affinity label for ribulose bisphosphate carboxylase/oxygenase from Rhodospirillum rubrum.

  1982cites this paper
• Synthesis of 1-chloro-2-oxohexanol 6-phosphate, a covalent active-site reagent for phosphoglucose isomerase.

  1981cites this paper
• Comparative inactivation and inhibition of the anomerase and isomerase activities of phosphoglucose isomerase

  1981cites this paper
• Affinity labeling and characterization of the active site histidine of glucosephosphate isomerase. Sequence homology with triosephosphate isomerase.

  1980cites this paper
• Paracatalytic modification of aldolase: a side reaction of the catalytic cycle resulting in irreversible blocking of two active-site lysyl residues.

  1979cites this paper
• The proton exchange of the pro-S hydrogen atom at C-1 in dihydroxyacetone phosphate and D-fructose 1,6-bisphosphate catalysed by class-I and class-II aldolases.

  1978cites this paper
• Affinity labeling of the pyridoxal phosphate binding site of the beta2 subunit of Escherichia coli tryptophan synthase.

  1978cites this paper
• Identification of essential lysyl and cysteinyl residues in spinach ribulosebisphosphate carboxylase/oxygenase modified by the affinity label N-bromoacetylethanolamine phosphate.

  1978cites this paper
• Attempts to apply affinity labeling techniques to ribulose bisphosphate carboxylase/oxygenase.

  1978cites this paper
• Rabbit muscle aldolase catalyzed proton exchange of hydroxyacetone phosphate with solvent.

  1977cites this paper
• Metabolism of D-fructose.

  1977cites this paper
• [46] Active-site-directed reagents of glycolytic enzymes

  1977cites this paper
• Detection of haloacetyl and haloketone affinity labels on paper chromatograms.

  1977cites this paper
• Inactivation of ribulosebisphosphate carboxylase/oxygenase from spinach with the affinity label N-bromoacetylethanolamine phosphate.

  1977cites this paper
• Affinity labelling of tRNA nucleotidyltransferase from baker's yeast with tRNAPhe modified on the 3'-terminus.

  1976cites this paper
• Affinity labeling of a previously undetected essential lysyl residue in class I fructose bisphosphate aldolase.

  1976cites this paper
• Detection of an essential sulfhydryl group in phosphoglycerate mutase with an affinity-labeling reagent.

  1976cites this paper
• Chloroplast and cytoplasmic enzymes: Subunit structure of pea leaf aldolases

  1975cites this paper
• Chloroplast and cytoplasmic enzymes 1, 2, 3. Subunit structure of pea leaf aldolases.

  1975cites this paper
• Chemical studies of enzyme active sites.

  1975cites this paper
• Identification of the histidyl residue of rabbit muscle aldolase alkylated by N-bromoacetylethanolamine phosphate.

  1974cites this paper
• The preparation of matrix-bound proteases and their use in the hydrolysis of proteins.

  1974cites this paper