Chiou S H, Pan F M, Peng H W, Chao Y K, Chang W C
Institute of Biochemical Sciences, National Taiwan University, Institute of Biological Chemistry, Academia Sinica, Taipei, Taiwan.
Biochem Biophys Res Commun. 1998 Nov 18;252(2):412-9. doi: 10.1006/bbrc.1998.9657.
gammaS-Crystallin from catfish eye lenses, formerly designated betas-crystallin in mammalian lenses, is structurally characterized in this study by cDNA cloning and sequencing. To facilitate sequence characterization of gammaS-crystallin with structural properties lying between beta- and gamma-crystallins, a cDNA mixture was constructed from the poly(A)+ mRNA isolated from catfish eye lenses, and amplification by polymerase chain reaction (PCR) was carried out to obtain nucleotide segments encoding multiple gammaS-crystallin isoforms. Sequencing several positive clones revealed that at least two distinct isoforms exist in the gammaS-crystallin class of this teleostean fish, similar to the authentic gamma-crystallin family characterized previously in species of the piscine class. Comparison of protein sequences encoded by two representative catfish gammaS1 and gammaS2 cDNAs with the published sequences of beta-, gamma-, and gammaS-crystallins from shark, carp, bullfrog, bovine, and human lenses indicates that there is about 20-50% sequence homology between catfish gammaS-crystallins and various members of the related beta/gamma-crystallin superfamily from different evolutionary classes, with a higher sequence similarity being found between catfish gammaS- and mammalian gamma-crystallins than between catfish gammaS- and bovine or carp gammaS-crystallins. Phylogenetic trees constructed on the basis of the nucleotide and protein sequence divergence among various beta-, gamma-, and gammaS-crystallins corroborate the closer relatedness of catfish gammaS- to authentic gamma-crystallin than to bovine and carp gammaS-crystallins. The results suggest that evolution of catfish gammaS-crystallins follows a different path from that of bovine and carp gammaS-crystallins and may represent a more ancient offshoot from the ancestral gamma/gammaS coding gene than carp and bovine gammaS-crystallins.
鲶鱼晶状体中的γS-晶体蛋白,以前在哺乳动物晶状体中被指定为βS-晶体蛋白,本研究通过cDNA克隆和测序对其结构进行了表征。为了便于对具有介于β-和γ-晶体蛋白之间结构特性的γS-晶体蛋白进行序列表征,从鲶鱼晶状体中分离的poly(A)+ mRNA构建了一个cDNA混合物,并通过聚合酶链反应(PCR)进行扩增,以获得编码多种γS-晶体蛋白异构体的核苷酸片段。对几个阳性克隆进行测序后发现,这种硬骨鱼的γS-晶体蛋白类别中至少存在两种不同的异构体,这与之前在鱼类物种中表征的正宗γ-晶体蛋白家族相似。将两个代表性的鲶鱼γS1和γS2 cDNA编码的蛋白质序列与已发表的鲨鱼、鲤鱼、牛蛙、牛和人晶状体的β-、γ-和γS-晶体蛋白序列进行比较,结果表明鲶鱼γS-晶体蛋白与来自不同进化类别的相关β/γ-晶体蛋白超家族的各种成员之间存在约20-50%的序列同源性,鲶鱼γS-与哺乳动物γ-晶体蛋白之间的序列相似性高于鲶鱼γS-与牛或鲤鱼γS-晶体蛋白之间的相似性。基于各种β-、γ-和γS-晶体蛋白之间的核苷酸和蛋白质序列差异构建的系统发育树证实,鲶鱼γS-与正宗γ-晶体蛋白的亲缘关系比与牛和鲤鱼γS-晶体蛋白更密切。结果表明,鲶鱼γS-晶体蛋白的进化与牛和鲤鱼γS-晶体蛋白的进化路径不同,可能代表了比鲤鱼和牛γS-晶体蛋白更古老的从祖先γ/γS编码基因分支出来的产物。