Matsubara Y, Kraus J P, Ozasa H, Glassberg R, Finocchiaro G, Ikeda Y, Mole J, Rosenberg L E, Tanaka K
J Biol Chem. 1987 Jul 25;262(21):10104-8.
cDNA encoding the precursor of rat liver medium chain acyl-CoA dehydrogenase (EC 1.3.99.3) was cloned and sequenced. The longest cDNA insert isolated was 1866 bases in length. This cDNA encodes the entire protein of 421-amino acids including a 25-amino acid leader peptide and a 396-amino acid mature polypeptide. The identity of the medium chain acyl-CoA dehydrogenase clone was confirmed by matching the amino acid sequence predicted from the cDNA to the NH2-terminal and nine internal tryptic peptide sequences derived from pure rat liver medium chain acyl-CoA dehydrogenase. The calculated molecular masses of the precursor medium chain acyl-CoA dehydrogenase, the mature medium chain acyl-CoA dehydrogenase, and the leader peptide are 46,600, 43,700, and 2,900 daltons, respectively. The leader peptide contains five basic amino acids and only one acidic amino acid; thus, it is positively charged, overall. Cysteine residues are unevenly distributed in the mature portion of the protein; five of six are found within the NH2-terminal half of the polypeptide. Comparison of medium chain acyl-CoA dehydrogenase sequence to other flavoproteins and enzymes which act on coenzyme A ester substrates did not lead to unambiguous identification of a possible FAD-binding site nor a coenzyme A-binding domain. The sequencing of other homologous acyl-CoA dehydrogenases will be informative in this regard.
编码大鼠肝脏中链酰基辅酶A脱氢酶（EC 1.3.99.3）前体的cDNA被克隆并测序。分离得到的最长cDNA插入片段长度为1866个碱基。该cDNA编码由421个氨基酸组成的完整蛋白质，包括一个25个氨基酸的前导肽和一个396个氨基酸的成熟多肽。通过将从cDNA预测的氨基酸序列与源自纯大鼠肝脏中链酰基辅酶A脱氢酶的NH2末端和九个内部胰蛋白酶肽序列进行匹配，确认了中链酰基辅酶A脱氢酶克隆的身份。前体中链酰基辅酶A脱氢酶、成熟中链酰基辅酶A脱氢酶和前导肽的计算分子量分别为46,600、43,700和2,900道尔顿。前导肽含有五个碱性氨基酸和仅一个酸性氨基酸；因此，总体上它带正电荷。半胱氨酸残基在蛋白质的成熟部分分布不均；六个中的五个位于多肽的NH2末端一半内。将中链酰基辅酶A脱氢酶序列与其他黄素蛋白和作用于辅酶A酯底物的酶进行比较，未明确鉴定出可能的FAD结合位点或辅酶A结合结构域。在这方面，其他同源酰基辅酶A脱氢酶的测序将提供信息。
