Reig J A, Kornblihtt A R, Flawiá M M, Torres H N
Biochem J. 1982 Oct 1;207(1):43-9. doi: 10.1042/bj2070043.
A soluble form of adenylate cyclase was extracted from mycelia of Neurospora crassa wild-type strains. This enzyme activity was purified by chromatography on hexyl-amino-Sepharose, agarose and Blue Sepharose and preparative polyacrylamide-gel electrophoresis. On sodium dodecyl sulphate/polyacrylamide-gel electrophoresis, peak fractions from the later purification steps showed a main polypeptide band with an apparent molecular weight of about 66 000. The following hydrodynamic and molecular parameters were established for the Neurospora soluble adenylate cyclase activity: sedimentation coefficient, 6.25 S; Stokes radius, 7.3 nm; partial specific volume, 0.74 ml/g; molecular weight, 202 000; frictional ratio, 1.65. The isoelectric point of this enzyme activity was 4.65. The enzyme was not activated by GTP, [beta gamma-imido]GTP, fluoride or cholera toxin.
从粗糙脉孢菌野生型菌株的菌丝体中提取出一种可溶性腺苷酸环化酶。该酶活性通过在己基氨基琼脂糖凝胶、琼脂糖凝胶和蓝色琼脂糖凝胶上进行层析以及制备性聚丙烯酰胺凝胶电泳进行纯化。在十二烷基硫酸钠/聚丙烯酰胺凝胶电泳中，后期纯化步骤的峰值馏分显示出一条主要的多肽带，其表观分子量约为66000。针对粗糙脉孢菌可溶性腺苷酸环化酶活性确定了以下流体动力学和分子参数：沉降系数为6.25 S；斯托克斯半径为7.3 nm；比容为0.74 ml/g；分子量为202000；摩擦比为1.65。该酶活性的等电点为4.65。该酶不受GTP、[βγ-亚氨基]GTP、氟化物或霍乱毒素的激活。
