Nishihira J, Ishibashi T, Sakai M, Nishi S, Kondo H, Makita A
Department of Biochemistry, Hokkaido University, Sapporo, Japan.
Biochem Biophys Res Commun. 1992 Nov 30;189(1):197-205. doi: 10.1016/0006-291x(92)91544-z.
Glutathione S-transferase P (GST-P) bound a series of endogenous fatty acids (C12-C18). To clarify the function and the binding site of the fatty acids, interaction between fatty acids and GST-P was investigated by using 12-(9-anthroyloxy) stearic acid conjugated with Woodward's reagent K. The fluorescence-conjugated fatty acid noncompetitively inhibited GST activity. After GST-P was covalently labeled with the fatty acid, the enzyme was digested with Lysyl Endopeptidase. From the peptide mapping, a single fluorescence-labeled peptide was obtained. By the sequence analysis, the peptide binding fatty acid was determined as the residues of 141-188 from the amino terminus.
谷胱甘肽S-转移酶P（GST-P）能结合一系列内源性脂肪酸（C12 - C18）。为阐明脂肪酸的功能及结合位点，通过使用与伍德沃德试剂K偶联的12 -（9 - 蒽氧基）硬脂酸研究了脂肪酸与GST-P之间的相互作用。荧光偶联脂肪酸非竞争性抑制GST活性。GST-P用脂肪酸共价标记后，用赖氨酰内肽酶消化该酶。通过肽图分析，得到了一个单一的荧光标记肽段。通过序列分析，确定结合脂肪酸的肽段为从氨基末端起的141 - 188位残基。
