Ungpakorn R, Holdom M D, Hamilton A J, Hay R J
Dermatology Unit, St John's Institute of Dermatology, Guy's Hospital, London, UK.
Clin Exp Dermatol. 1996 May;21(3):190-6. doi: 10.1111/j.1365-2230.1996.tb00060.x.
Cell homogenization, isoelectric focusing and gel filtration FPLC have been used to purify a superoxide dismutase (SOD) from the dermatophyte Trichophyton mentagrophytes var. interdigitale (T. interdigitale). N-terminal amino acid sequencing identified this enzyme as a Cu,ZnSOD, with a pH of 5.1, a reduced molecular mass of 18 kDa, and a non-reduced molecular mass of 59 kDa. SOD activity was detectable in culture filtrates, as early as the mid-log phase of growth. The known Cu,Zn inhibitor potassium cyanide caused some inhibition of the purified enzyme, whereas the inhibitors sodium azide, guanidinium hydrochloride, EDTA and chloroform/ethanol had no discernible effect. The T. interdigitale SOD was pH insensitive in the range 7.0-10.5 and remained active after prolonged incubation at 50 degrees C. The purification and characterization of this enzyme represents the first step in determining whether SOD plays any part in protecting T. interdigitale from free radicals generated by the oxidative burst of immune effector cells.
已使用细胞匀浆、等电聚焦和凝胶过滤快速蛋白质液相色谱法从皮肤癣菌须癣毛癣菌指间变种（T. interdigitale）中纯化超氧化物歧化酶（SOD）。N端氨基酸测序确定该酶为铜锌超氧化物歧化酶，其pH值为5.1，还原分子量为18 kDa，非还原分子量为59 kDa。早在生长对数中期，在培养滤液中就能检测到超氧化物歧化酶活性。已知的铜锌抑制剂氰化钾对纯化后的酶有一定抑制作用，而抑制剂叠氮化钠、盐酸胍、乙二胺四乙酸和氯仿/乙醇则没有明显影响。指间毛癣菌超氧化物歧化酶在7.0 - 10.5范围内对pH不敏感，在50℃长时间孵育后仍保持活性。该酶的纯化和特性鉴定是确定超氧化物歧化酶是否在保护指间毛癣菌免受免疫效应细胞氧化爆发产生的自由基影响方面发挥任何作用的第一步。
