Kikuchi K, Yamashita M, Watabe S, Aida K
Laboratory of Fish Physiology, Faculty of Agriculture, University of Tokyo, Japan.
J Biol Chem. 1995 Jul 21;270(29):17087-92. doi: 10.1074/jbc.270.29.17087.
cDNAs encoding a warm temperature acclimation-related protein (Wap65) were cloned from the muscle and hepatopancreas cDNA libraries of the warm temperature-acclimated goldfish Carassius auratus, and their nucleotide sequences containing 5'- and 3'-noncoding regions together with their polyadenylation signal were determined. The deduced amino acid sequence of Wap65 was 31% homologous to rat hemopexin. However, goldfish Wap65 lacked a few possible glycosylation sites and presumed functional histidine residues, implying that it may have different functions from hemopexin. Wap65 contained a leader peptide of 30 amino acids and a mature protein region of 415 amino acids. Southern blot analysis demonstrated that the protein is expressed by a single copy gene in the goldfish haploid genome. In RNA blot analysis using isolated cDNA clones, a single transcript of about 2.0 kilobases was detected in the hepatopancreas but not in brain, muscle, or hemocytes. The abundancy of this transcript markedly increased in the hepatopancreas as a result of warm temperature acclimation. Electrophoretic analysis of plasma proteins revealed a good correlation of plasma Wap65 levels to those of the corresponding transcript in the hepatopancreas, suggesting that serum Wap65 concentrations are regulated mainly by transcript levels in the hepatopancreas via the secretion process.
从经暖温驯化的金鱼（Carassius auratus）的肌肉和肝胰腺cDNA文库中克隆出编码一种与暖温驯化相关蛋白（Wap65）的cDNA，并测定了其包含5'-和3'-非编码区以及多聚腺苷酸化信号的核苷酸序列。推导的Wap65氨基酸序列与大鼠血红素结合蛋白有31%的同源性。然而，金鱼Wap65缺少一些可能的糖基化位点和推测的功能性组氨酸残基，这意味着它可能具有与血红素结合蛋白不同的功能。Wap65包含一个30个氨基酸的前导肽和一个415个氨基酸的成熟蛋白区域。Southern印迹分析表明，该蛋白由金鱼单倍体基因组中的单拷贝基因表达。在使用分离的cDNA克隆进行的RNA印迹分析中，在肝胰腺中检测到一个约2.0千碱基的单一转录本，而在脑、肌肉或血细胞中未检测到。由于暖温驯化，该转录本在肝胰腺中的丰度显著增加。血浆蛋白的电泳分析显示，血浆Wap65水平与肝胰腺中相应转录本的水平具有良好的相关性，这表明血清Wap65浓度主要通过分泌过程由肝胰腺中的转录本水平调节。
