Hagiya M, Francavilla A, Polimeno L, Ihara I, Sakai H, Seki T, Shimonishi M, Porter K A, Starzl T E
Pharmaceuticals Research Center, Toyobo Co., Ltd, Shiga, Japan.
Proc Natl Acad Sci U S A. 1994 Aug 16;91(17):8142-6. doi: 10.1073/pnas.91.17.8142.
A full-length cDNA clone encoding a purified augmenter of liver regeneration (ALR) factor prepared from the cytosol of weanling rat livers was isolated. The 1.2-kb cDNA included a 299-bp 5' untranslated region, a 375-bp coding region, and a 550-bp 3' untranslated region. It encoded a protein consisting of 125 amino acids. The molecular weight of ALR calculated from the cDNA was 15,081, which is consistent with the size estimated by SDS/PAGE under reducing conditions. The molecular weight of the purified native ALR estimated by SDS/PAGE under nonreducing conditions was approximately 30,000; thus ALR apparently has a homodimeric structure. The recombinant ALR produced by expression of the cDNA in COS cells was tested in vivo in the canine Eck fistula model and found to have potency equivalent to the purified native ALR. The 125-aa sequence deduced from the rat ALR cDNA shows 50% homology to the amino acid sequence of the gene for oxidative phosphorylation and vegetative growth in the yeast Saccharomyces cerevisiae.
从断奶大鼠肝脏胞质溶胶中制备的编码纯化的肝再生增强因子(ALR)的全长cDNA克隆被分离出来。这个1.2kb的cDNA包含一个299bp的5'非翻译区、一个375bp的编码区和一个550bp的3'非翻译区。它编码一种由125个氨基酸组成的蛋白质。根据cDNA计算的ALR分子量为15081,这与在还原条件下通过SDS/PAGE估计的大小一致。在非还原条件下通过SDS/PAGE估计的纯化天然ALR的分子量约为30000;因此,ALR显然具有同二聚体结构。通过在COS细胞中表达cDNA产生的重组ALR在犬Eck瘘模型中进行了体内测试,发现其效力与纯化的天然ALR相当。从大鼠ALR cDNA推导的125个氨基酸序列与酿酒酵母中氧化磷酸化和营养生长基因的氨基酸序列具有50%的同源性。