Feldberg R S, Iannitti D A, Cochrane D E
Department of Biology, Tufts University, Medford, MA 02155.
Biochem J. 1988 Jan 1;249(1):297-300. doi: 10.1042/bj2490297.
A procedure for obtaining rat mast-cell histidine decarboxylase in greater than 50% yield in cell-free extracts was developed. The enzyme was found in the supernatant fractions from a 3,500 g and a 105,000 g centrifugation step and was demonstrated to be sensitive to inhibition by alpha-fluoromethylhistidine but not by phenylalanine. Although the enzyme shows a half-life of only 3 h in cell-free extract, the initial high recovery of activity allowed for active-site labelling of the enzyme by [3H]histidine and NaBH4. Labelled protein migrated on non-denaturing polyacrylamide-gradient-gel electrophoresis as a 55,000 Da species.
已开发出一种程序，可在无细胞提取物中以超过50%的产率获得大鼠肥大细胞组氨酸脱羧酶。该酶存在于3500g和105000g离心步骤的上清液部分，并且已证明它对α-氟甲基组氨酸敏感，但对苯丙氨酸不敏感。尽管该酶在无细胞提取物中的半衰期仅为3小时，但活性的初始高回收率使得能够用[3H]组氨酸和硼氢化钠对该酶进行活性位点标记。标记的蛋白质在非变性聚丙烯酰胺梯度凝胶电泳上以55000Da的条带迁移。
