Magnuson M A, Morioka H, Tecce M F, Nikodem V M
J Biol Chem. 1986 Jan 25;261(3):1183-6.
The nucleotide sequence of the mRNA for malic enzyme ((S)-malate NADP+ oxidoreductase (oxaloacetate-decarboxylating, EC 1.1.1.40) from rat liver was determined from three overlapping cDNA clones. Together, these clones contain 2078 nucleotides complementary to rat liver malic enzyme mRNA. The single open reading frame of 1761 nucleotides codes for a 585-amino acid polypeptide with a calculated molecular mass of about 65,460 daltons. The cloned cDNAs contain the complete 3'-noncoding region of 301 nucleotides for the major mRNA species of rat liver and 16 nucleotides of the 5'-noncoding region. Amino acid sequences of seven tryptic peptides (67 amino acids) from the purified protein are distributed through the single open reading frame and show excellent correspondence with the translated nucleotide sequence. The putative NADP-binding site for malic enzyme was identified by amino acid sequence homology with the NADP-binding site of the enoyl reductase domain of fatty acid synthetase.
通过三个重叠的cDNA克隆确定了大鼠肝脏苹果酸酶((S)-苹果酸NADP +氧化还原酶(草酰乙酸脱羧,EC 1.1.1.40))mRNA的核苷酸序列。这些克隆一起包含2078个与大鼠肝脏苹果酸酶mRNA互补的核苷酸。1761个核苷酸的单一开放阅读框编码一个585个氨基酸的多肽,计算分子量约为65,460道尔顿。克隆的cDNA包含大鼠肝脏主要mRNA物种301个核苷酸的完整3'-非编码区和5'-非编码区的16个核苷酸。纯化蛋白的七个胰蛋白酶肽(67个氨基酸)的氨基酸序列分布在单一开放阅读框中,并与翻译的核苷酸序列显示出极好的对应性。通过与脂肪酸合成酶烯酰还原酶结构域的NADP结合位点的氨基酸序列同源性鉴定了苹果酸酶的推定NADP结合位点。