Gene/Protein Disease Symptom Drug Enzyme Compound
Pivot Concepts:   Target Concepts:
Query: EC:2.7.1.1 (hexokinase)
 5,274 document(s) hit in 31,850,051 MEDLINE articles (0.00 seconds)

In yeast hexokinase B, two thiols per monomer appeared to be essential when enzymic inactivation was produced by the concurrent alkylation of both of them, by several reagents including the affinity reagent N-bromoacetyl-2-D-galactosamine. However, it is shown that only one of these thiols is actually essential. Three of the four thiols present can be blocked by alkylation in the presence of a substrate in appropriate conditions, without loss of enzymic activity. Subsequently, in the absence of substrate, the affinity reagent reacts at the one remaining thiol, with complete inactivation. The same behavior can be obtained by reaction with iodoacetamide or by the formation of the -SCN group. The affinity reagent inactivates hexokinase B faster than does the isomeric glycosidic compound (glycosides being nonsubstrates), although the latter has twice the reactivity of the former toward glutathione. The reactions with alkylating agents, with or without substrate present, are used to classify the four thiols in the monomer. The temperature dependence of the alkylation of the essential thiol provides evidence for a transition in the molecule at about 31 degrees C. The inactive monomer containing the -SCN group can regenerate, by thiolysis, active enzyme with the thiol free. It can also perform an intramolecular cleavage of the chain. The latter reaction was used to locate the essential cysteine residue in the chain, at 80% of the length from the N terminus.
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PMID:Evidence for a single essential thiol in the yeast hexokinase molecule. 33 26

The enzymatic activity of hexokinase (ATP : D-hexose 6-phosphotransferase, EC 2.7.1.1) decreased rapidly when the enzyme was exposed to the lactoperoxidase antimicrobial system (consisting of lactoperoxidase, H2O2 and SCN-). Inactivation did not begin until the reaction of one sulfhydryl group per hexokinase monomer was completed. Loss of enzyme activity accompanied the reaction of at least one additional sulfhydryl group per monomer. Covalent incorporation of 14C-labeled SCN- into hexokinase increased as the inactivation reaction progressed. The rate of the hexokinase activity loss dependent on temperature, pH and the presence of glucose and phosphate ion. When H2O2 and SCN- were applied to a Sepharose column bearing covalently attached lactoperoxidase, the column eluate inactivated hexokinase. This demonstrated that the lactoperoxidase molecule itself need not be in contact with hexokinase in order to catalyze hexokinase inactivation. The sulfhydryl-reactive oxidation product of SCN- which is generated by the column is sufficient. The results are consistent with a two-stage reaction in which the exposed, non-essential sulfhydryl groups on the hexokinase molecule react first to produce an enzymatically active but unstable form of hexokinase. This modified form of hexokinase then undergoes a spontaneous, temperature-dependent structural change, which allows reaction of previously shielded, essential sulfhydryl groups. The phenomenon described here suggests a possible mechanism for the antimicrobial effects of the lactoperoxidase system.
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PMID:Lactoperoxidase-catalyzed inactivation of hexokinase. 724 2