Peeters R A, Veerkamp J H, Geurts van Kessel A, Kanda T, Ono T
Department of Biochemistry, University of Nijmegen, The Netherlands.
Biochem J. 1991 May 15;276 ( Pt 1)(Pt 1):203-7. doi: 10.1042/bj2760203.
A cDNA clone for the human skeletal-muscle fatty-acid-binding protein (FABP) was isolated by screening of a human adult muscle lambda gt11 expression library with an anti-(muscle FABP) serum. The identify of the clone was confirmed by transcription/translation in vitro in plasmid pSP6.5, followed by immunoprecipitation. The nucleotide sequence of the 551 bp cDNA insert showed an open reading frame of 399 nucleotides, coding for a protein of 133 amino acid residues with a calculated molecular mass of 14858 Da and a pI of 4.94. Only one cysteine residue was found, at position 125. Peptide sequence analyses of human skeletal-muscle and heart FABP, after carbamoylmethylation and lysyl endopeptidase digestion followed by automatic Edman degradation, showed that both proteins are identical. No evidence was found for the existence of isoproteins in muscle. The chromosomal localization of the human muscle FABP gene was determined by analysing 31 human x rodent somatic-cell hybrid lines. The human muscle FABP gene could be assigned to chromosome 1pter-q31.
通过用抗(肌肉脂肪酸结合蛋白)血清筛选人成人肌肉λgt11表达文库,分离出了人骨骼肌脂肪酸结合蛋白(FABP)的cDNA克隆。通过在质粒pSP6.5中进行体外转录/翻译,随后进行免疫沉淀,证实了该克隆的身份。551bp cDNA插入片段的核苷酸序列显示有一个399个核苷酸的开放阅读框,编码一个由133个氨基酸残基组成的蛋白质,计算分子量为14858Da,pI为4.94。仅在第125位发现一个半胱氨酸残基。对人骨骼肌和心脏FABP进行氨甲酰甲基化和赖氨酰内肽酶消化,然后进行自动Edman降解后的肽序列分析表明,这两种蛋白质是相同的。未发现肌肉中存在同工蛋白的证据。通过分析31个人-啮齿动物体细胞杂种系,确定了人肌肉FABP基因的染色体定位。人肌肉FABP基因可定位于染色体1pter-q31。