Photoaffinity labelling of carnitine acetyltransferase with S-(p-azidophenacyl)thiocarnitine.
Photoaffinity labelling of carnitine acetyltransferase with S-(p-azidophenacyl)thiocarnitine.
复制标题
用 S-（对叠氮苯甲酰基）硫代肉碱对肉碱乙酰转移酶进行光亲和标记。
DOI：
10.1042/bj2370533
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发表时间：
1986
期刊：
影响因子：
--
通讯作者：
Barden,RE
中科院分区：
文献类型：
--
作者：
Mauro,JM;Lewis,RV;Barden,RE
A photolabile reagent, p-azidophenacyl-DL-thiocarnitine, was synthesized and tested as a photoaffinity label for carnitine acetyltransferase (EC 2.3.1.7) from pigeon breast. p-Azidophenacyl-DL-thiocarnitine is an active-site-directed reagent for this acetyltransferase, since it is a competitive inhibitor (Ki 10 microM) versus carnitine. U.v. irradiation of a mixture of p-azidophenacyl-DL-thiocarnitine and enzyme produces irreversible inhibition. Acetyl-DL-carnitine protects the enzyme from inhibition by photoactivated p-azidophenacyl-DL-thiocarnitine. In the presence of 30 mM-2-mercaptoethanol as a scavenger, the relationship between loss of activity and photoincorporation of reagent suggests that one molecule of reagent is incorporated per molecule of inhibited enzyme. However, peptide maps of enzyme labelled with p-azidophenacyl[14C]thiocarnitine indicate that several (about six) tryptic peptides (of a possible 60-65) are modified. The presence of 5 mM-acetyl-DL-carnitine significantly decreases the incorporation of reagent in each labelled tryptic peptide.
