title: [Endo-beta-1,3-glucanase using affinity inhibition].
abstract: Synthetic 2',3'-epoxypropyl-1-thio-beta-D-glucopyranoside selectively modifies a catalytically essential nucleophylic group in the active site of beta-1,3-glucanase LIV from the marine mollusc Spisula sachalinensis, the inactivation being as high as 95%. The properties of native and epoxypropylthioglucopyranoside-inhibited glucanase LIV were compared, using UV-spectroscopy, SDS polyacrylamide gel electrophoresis and isoelectrofocusing. It was found that the addition of laminarine and laminarioligosaccharides to a solution of the inhibited enzyme induces UV-differential spectra typical for the tryptophanyl residue involved in the formation of the enzyme-inhibitor-substrate complex. The glucone-1,5-lactone does not produce such spectrum. It was shown that epoxypropylthioglucopyranoside protects the accessible tryptophanyl residues in the enzyme active center against the oxidation by N-bromosuccinimide.
mesh_terms: D000345:Affinity Labels / Q000494:pharmacology*; D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel; D004693:Glucan Endo-1,3-beta-D-Glucosidase / Q000037:antagonists & inhibitors*; D006026:Glycoside Hydrolases / Q000037:antagonists & inhibitors*; D007700:Kinetics; D008974:Mollusca / Q000201:enzymology*; D013056:Spectrophotometry, Ultraviolet; D013865:Thioglycosides / Q000494:pharmacology*
chemical_list: D000345:Affinity Labels; D013865:Thioglycosides; C039912:2',3'-epoxypropyl-1-thioglucoside; D006026:Glycoside Hydrolases; D004693:Glucan Endo-1,3-beta-D-Glucosidase
vernacular_title: Issledovanie éndo-beta-1,3-gliukanazy metodom affinnogo ingibirovaniia.