用于鉴定光敏真菌粗糙脉孢菌细胞内游离黄素种类的方法。

Method for identification of intracellular free flavin species in the photosensitive fungus Neurospora crassa.

作者信息

Scheideler L, Ninnemann H

机构信息

Institut für Chemische Pflanzenphysiologie, Universität Tübingen, Federal Republic of Germany.

出版信息

Anal Biochem. 1988 Aug 15;173(1):106-10. doi: 10.1016/0003-2697(88)90166-2.

DOI:10.1016/0003-2697(88)90166-2

PMID:2973261

Abstract

Establishing the relative intracellular proportions of flavins in Neurospora crassa (and in other organisms) in vivo may be hampered by degradation of flavins after homogenization of the cells. The system described here allows separation and identification of intracellular free and bound flavins under conditions restrictive for the FAD-degrading enzyme(s). A "protective buffer" containing 0.1 M citrate adjusted to pH 4.0 with K2HPO4, 5 mM ATP, and 0.5 mM EDTA prevents FAD from rapid enzymatic cleavage in crude cell lysates of the Neurospora crassa mutant "slime."

摘要

细胞匀浆后黄素的降解可能会妨碍确定粗糙脉孢菌（以及其他生物体）体内黄素的相对细胞内比例。本文所述的系统能够在限制FAD降解酶的条件下分离并鉴定细胞内游离和结合的黄素。一种“保护缓冲液”，其含有用K2HPO4调节至pH 4.0的0.1 M柠檬酸盐、5 mM ATP和0.5 mM EDTA，可防止粗糙脉孢菌突变体“黏液”的粗细胞裂解物中FAD被快速酶解。