Ha H, Hajek P, Bedwell D M, Burrows P D
Department of Microbiology, University of Alabama, Birmingham 35294.
J Biol Chem. 1993 Jun 5;268(16):12143-9.
Pores formed in the outer membrane of mitochondria by mitochondrial porin make it permeable to water-soluble metabolites smaller than approximately 5 kDa. We have isolated a full-length cDNA encoding a human porin. This probe detects a single approximately 1.5-kilobase mRNA species on Northern blots, but multiple hybridizing bands on genomic Southern blots. The open reading frame predicts a 38.1-kDa protein with a pI of 6.59 that is homologous but not identical to a previously reported protein sequence of a 31-kDa porin isolated from human lymphocytes (porin 31HL). The most striking difference between the two porins is that the sequence predicted by the cDNA is longer than the 31HL porin by 27 amino acids at the amino terminus and 38 amino acids at the carboxyl terminus. The porin cDNA directs the in vitro translation of two protein species of approximately 32 and approximately 36 kDa, which appear to result from alternate usage of AUG initiation codons. The 32-kDa protein is the predominant species imported into both rat and yeast mitochondria in vitro. Taken together, these results suggest that multiple porin proteins can be expressed in humans. Additionally, a porin consensus protein sequence has been identified that is conserved in eukaryotic organisms as diverse as yeast and man.
线粒体孔蛋白在线粒体外膜上形成的孔道,使线粒体对分子量小于约5 kDa的水溶性代谢产物具有通透性。我们分离出了编码人孔蛋白的全长cDNA。该探针在Northern印迹上检测到一条约1.5千碱基的mRNA条带,但在基因组Southern印迹上检测到多条杂交带。开放阅读框预测的蛋白质分子量为38.1 kDa,pI为6.59,与先前报道的从人淋巴细胞中分离出的31 kDa孔蛋白(孔蛋白31HL)的蛋白质序列同源但不相同。这两种孔蛋白最显著的差异在于,cDNA预测的序列在氨基末端比31HL孔蛋白长27个氨基酸,在羧基末端长38个氨基酸。孔蛋白cDNA指导体外翻译出两种分子量分别约为32 kDa和约36 kDa的蛋白质,这似乎是由于AUG起始密码子的交替使用所致。在体外,32 kDa的蛋白质是导入大鼠和酵母线粒体的主要蛋白质。综上所述,这些结果表明人可以表达多种孔蛋白。此外,还鉴定出了一种在酵母和人等多种真核生物中保守的孔蛋白共有蛋白质序列。