Kondo H, Kawazoe I, Nakaya M, Kikuchi K, Aida K, Watabe S
Division of Aquatic Bioscience, Graduate School of Agricultural and Life Sciences, University of Tokyo, Bunkyo, 113-8657, Tokyo, Japan.
Biochim Biophys Acta. 2001 Mar 30;1531(1-2):132-42. doi: 10.1016/s1388-1981(01)00099-3.
cDNAs encoding major plasma apolipoproteins (apo) were cloned from the eel Anguilla japonica liver and their nucleotide sequences determined. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed that eel lipoproteins contain apolipoproteins of 28 kDa and 14 kDa as major components. Each of the two apolipoproteins showed two isoforms having different isoelectric points as demonstrated by two-dimensional electrophoresis. The two 28 kDa components had different N-terminal amino acid sequences, whereas the two 14 kDa components had an identical one. Then cDNA clones encoding these apolipoproteins were isolated from a cDNA library constructed from the eel liver. An acidic 28 kDa component (28 kDa-1) consisted of 259 amino acids including a putative signal peptide of 27 residues, whereas a basic 28 kDa component (28 kDa-2) was composed of 260 amino acids containing a putative signal peptide of 23 residues. The tandem repeating units, which are characteristic of apolipoproteins, for 28 kDa-1 showed 27.8% identity to that of porcine apoA-IV, although mammalian apoA-IV is about 40 kDa and much larger than 28 kDa-1. However, the repeating units of 28 kDa-2 showed 52.5% identity to that of Atlantic salmon apoA-I. The 14 kDa apolipoprotein consisted of 142 amino acids containing a putative signal peptide of 20 residues. It has a novel sequence differing from apolipoproteins of other vertebrates. The transcriptional expressions of 28 kDa-1, 28 kDa-2, and 14 kDa components were all restricted to the liver, except for the transcripts of 28 kDa-2 which were also slightly expressed in the intestine.
从日本鳗鲡肝脏中克隆出编码主要血浆载脂蛋白(apo)的cDNA,并测定了它们的核苷酸序列。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳显示,鳗脂蛋白含有28 kDa和14 kDa的载脂蛋白作为主要成分。二维电泳表明,这两种载脂蛋白各自都有两种具有不同等电点的异构体。两种28 kDa的成分具有不同的N端氨基酸序列,而两种14 kDa的成分具有相同的N端氨基酸序列。然后从鳗肝脏构建的cDNA文库中分离出编码这些载脂蛋白的cDNA克隆。一种酸性28 kDa成分(28 kDa-1)由259个氨基酸组成,包括一个27个残基的假定信号肽,而一种碱性28 kDa成分(28 kDa-2)由260个氨基酸组成,含有一个23个残基的假定信号肽。28 kDa-1的串联重复单元是载脂蛋白的特征,与猪apoA-IV的串联重复单元有27.8%的同一性,尽管哺乳动物的apoA-IV约为40 kDa,比28 kDa-1大得多。然而,28 kDa-2的重复单元与大西洋鲑鱼apoA-I的重复单元有52.5%的同一性。14 kDa的载脂蛋白由142个氨基酸组成,含有一个20个残基的假定信号肽。它有一个与其他脊椎动物载脂蛋白不同的新序列。28 kDa-1、28 kDa-2和14 kDa成分的转录表达都局限于肝脏,除了28 kDa-2的转录本在肠道中也有轻微表达。