IDENTIFICATION OF A HIGHLY REACTIVE SULFHYDRYL-GROUP IN HUMAN PLACENTAL GLUTATHIONE TRANSFERASE BY A SITE-DIRECTED FLUORESCENT REAGENT

被引：51

作者：

LOBELLO, M ^{[1
]}

PETRUZZELLI, R ^{[1
]}

DESTEFANO, E ^{[1
]}

TENEDINI, C ^{[1
]}

BARRA, D ^{[1
]}

FEDERICI, G ^{[1
]}

机构：

[1] UNIV ROME LA SAPIENZA,DEPT BIOCHEM SCI,I-00185 ROME,ITALY

来源：

FEBS LETTERS | 1990年 / 263卷 / 02期

关键词：

Chemical modification; Fluorescent probe; Glutathione transferase; Sulphydryl group;

D O I：

10.1016/0014-5793(90)81421-J

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

A fluorescent maleimide derivative, N-(4-anilino-1-naphthyl) maleimide (ANM), a specific probe for thiol groups, reacted with human placental glutathione transferase (GST, EC 2.5.1.18), causing a complete inactivation of the enzyme in a few minutes. The modified enzyme was denatured, alkylated and digested with (L-1-tosylamide-2-phenylethyl chloromethyl ketone)-trypsin. The tryptic digest was analysed by HPLC and a fluorescent peptide was obtained. The sequence of this peptide allowed us, by a comparison with a well known primary structure, to assign the position 47 to the most reactive cysteine of GST enzyme. © 1990.

引用

页码：389 / 391

页数：3

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