IDENTIFICATION OF KONINGIC ACID (HEPTELIDIC ACID)-MODIFIED SITE IN RABBIT MUSCLE GLYCERALDEHYDE-3-PHOSPHATE DEHYDROGENASE
IDENTIFICATION OF KONINGIC ACID (HEPTELIDIC ACID)-MODIFIED SITE IN RABBIT MUSCLE GLYCERALDEHYDE-3-PHOSPHATE DEHYDROGENASE
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DOI：
10.1016/0167-4838(91)90058-8
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发表时间：
1991-04-08
期刊：
影响因子：
--
通讯作者：
ENDO, A
中科院分区：
文献类型：
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作者：
SAKAI, K;HASUMI, K;ENDO, A
倍半萜类抗生素柯宁酸是一种特殊的修饰3-磷酸甘油醛脱氢酶的药物(Sakai，K.，Hasumi，K.和Endo，A.(1988)Biochim)。生物群落。会刊952,297-303)。用胰酶消化[H-3]康宁酸标记的兔肌肉甘油醛-3-磷酸脱氢酶。反相高效液相色谱法显示，该标记仅与一种含有17个氨基酸残基的胰蛋白酶多肽相关。显微测序和快原子轰击质谱分析表明，该多肽具有Ile-Var-Ser-Asn-Ala-Ser-Cys-Thr-Thr-Asn-Cys-Leu-Ala-Pro-Leu-Ala-Lys.序列与其他物种的甘油醛-3-磷酸脱氢酶的氨基酸序列相比，该肽位于高度保守的区域，是该酶活性部位的一部分。猪肌肉酶中与Cys-149相对应的半胱氨酸残基被证明是该酶活性所必需的残基，是椰子酸修饰的部位。对康宁酸与L-半胱氨酸反应产物的结构分析表明，康宁酸的环氧化物与半胱氨酸残基的巯基反应生成硫醚。
The sesquiterpene antibiotic koningic acid (heptelidic acid) has been previously demonstrated to modify glyceraldehyde-3-phosphate dehydrogenase in specific manner, probably by binding to the sulfhydryl residue at the active site of the enzyme (Sakai, K., Hasumi, K. and Endo, A. (1988) Biochim. Biophys. Acta 952, 297-303). Rabbit muscle glyceraldehyde-3-phosphate dehydrogenase labeled with [H-3]koningic acid was digested with trypsin. Reverse-phase HPLC revealed that the label is associated exclusively with a tryptic peptide having 17 amino acid residues. Microsequencing and fast atom bombardment mass spectrometry demonstrated that the peptide has the sequence Ile-Var-Ser-Asn-Ala-Ser-Cys-Thr-Thr-Asn-Cys-Leu-Ala-Pro-Leu-Ala-Lys. In comparison to the amino acid sequence of glyceraldehyde-3-phosphate dehydrogenase from other species, this peptide is in a highly conserved region and is part of the active site of the enzyme. The cysteine residue corresponding to the Cys-149 in the pig muscle enzyme, which has been shown to be an essential residue for the enzyme activity, was shown to be the site modified by koningic acid. Structural analyses of the reaction product of koningic acid and L-cysteine suggested that the epoxide of koningic acid reacts with the sulfhydryl group of cysteine residue, resulting in a thioether.