Lambkin I, Hamilton A J, Hay R J
Dermatology Unit, Clinical Sciences Laboratory, Guys Hospital, London, UK.
Mycoses. 1994 Mar-Apr;37(3-4):85-92. doi: 10.1111/j.1439-0507.1994.tb00782.x.
An extracellular proteinase has been partially purified from culture filtrates of Trichophyton rubrum by ultrafiltration, isoelectric focusing and gel filtration chromatography. The enzyme has a non-reduced molecular weight of 235,000 by substrate SDS-PAGE. It has a pH optimum of 8.5 using azocasein and azoalbumin as substrates and a pI of 3.6-3.8. The metalloproteinase inhibitors EDTA and 1,10-phenanthroline, together with the chymotrypsin inhibitor chymostatin, strongly inhibited its activity. The serine proteinase inhibitors phenylmethanesulphonyl fluoride and diisopropylfluorophosphate showed weak inhibitory activity. The proteinase exhibited broad substrate activity against azocoll, azoalbumin, azocasein, laminin and fibronectin. It exhibited weak activity against elastin and keratin. Observations on the occurrence of this proteinase together with previously described lower molecular weight proteinases suggests that the former is the first to appear in minimal medium cultures. Freeze/thaw cycling of the partially purified 235,000 M(r) proteinase was found to generate low molecular weight proteinases, particularly at 53,000, 27,000 and 25,000 M(r), indicating that the latter may originate from the larger molecule.
通过超滤、等电聚焦和凝胶过滤色谱法，从红色毛癣菌的培养滤液中部分纯化了一种细胞外蛋白酶。用底物SDS-PAGE测定，该酶的非还原分子量为235,000。以偶氮酪蛋白和偶氮白蛋白为底物时，其最适pH为8.5，pI为3.6 - 3.8。金属蛋白酶抑制剂EDTA和1,10 - 菲咯啉以及糜蛋白酶抑制剂抑糜酶素强烈抑制其活性。丝氨酸蛋白酶抑制剂苯甲基磺酰氟和二异丙基氟磷酸表现出较弱的抑制活性。该蛋白酶对偶氮胶原、偶氮白蛋白、偶氮酪蛋白、层粘连蛋白和纤连蛋白具有广泛的底物活性。它对弹性蛋白和角蛋白的活性较弱。对这种蛋白酶以及先前描述的低分子量蛋白酶出现情况的观察表明，前者是在基本培养基培养物中最先出现的。发现部分纯化的235,000 M(r)蛋白酶经冻融循环会产生低分子量蛋白酶，特别是分子量为53,000、27,000和25,000 M(r)的蛋白酶，这表明后者可能起源于较大的分子。
