The activating system of chitin synthetase from Saccharomyces cerevisiae. Purification and properties of the activating factor.

摘要: The yeast proteinase that causes activation of the chitin synthetase zymogen has been purified by a procedure includes affinity chromatography on an agarose column to which proteinaceous inhibitor enzyme had covalently attached. yielded single band upon disc gel electrophoresis at pH 4.5 in presence urea. At same pH, but without urea, faint was detected coincidence with enzymatic activity, whereas 9.5, either absence or sodium dodecyl sulfate, no protein zone could be seen. From sedimentation and filtration data, molecular weight 44,000 estimated. active within wide range values, optimum between 6.5 AND 7. Titraton activity from required 1 mol inhibitor/mol enzyme. A similar result obtained phenylmethylsulfonyl fluoride, indication serine residue is for activity. exhibited hydrolytic several proteins esterolytic many synthetic substrates, including benzoylarginine ethyl ester acetyltyrosine ester.A comparison properties those known proteinases led conclusion synthestase activating factor identical previously designated as B (EC 3.4.22.9). This first time homogeneous preparation characterized.