Obokata J, Mikami K, Hayashida N, Nakamura M, Sugiura M
Department of Botany, Faculty of Science, Hokkaido University, Sapporo, Japan.
Plant Physiol. 1993 Aug;102(4):1259-67. doi: 10.1104/pp.102.4.1259.
The protein composition of photosystem I (PSI) was examined in Nicotiana spp. by high-resolution polyacrylamide gel electrophoresis, N-terminal amino acid sequencing, and immunoblot analysis. Five PSI proteins show polymorphism in an amphidiploid species, Nicotiana tabacum, but not in its ancestral diploid species, Nicotiana sylvestris and Nicotiana tomentosiformis. These Nicotiana spp. appear to have at least 18 PSI proteins per genome that range in molecular mass from 3 to 20 kD. They include the products of nuclear genes psaD, psaE, psaF, psaG, psaH, psaK, and psaL, the product of chloroplast gene psaC, N-terminally blocked proteins of 4.5 and 3.0 kD, and an unidentified protein of 12.5 kD. The psaD, psaF, psaH, and psaL products have two isoforms each that are distinguished by different mobilities in polyacrylamide gel electrophoresis, and the psaE product has four isoforms. The two isoforms of the psaD product have distinct amino acid sequences, indicating that they are encoded by different genes within the genome. Four isoforms of the psaE products can be classified into two groups by N-terminal amino acid sequence, indicating that at least two psaE genes are present in the genome. To examine whether the polymorphic nature of PSI is peculiar to Nicotiana spp., we carried out immunoblot analysis of the psaD and psaE products in isogenic lines of tomato (Lycopersicon esculentum), Arabidopsis thaliana, red bean (Vigna angularis), and corn (Zea mays). Two electrophoretically distinct isoforms were found for the psaD products of tomato, A. thaliana, and corn, and two isoforms of psaE products were detected in tomato, A. thaliana, and red bean. These results suggest that the nuclear-encoded subunits of PSI, except for the psaG and psaK products, generally have two isoforms.
通过高分辨率聚丙烯酰胺凝胶电泳、N 端氨基酸测序和免疫印迹分析,对烟草属植物的光系统 I(PSI)的蛋白质组成进行了研究。在异源四倍体物种烟草中,有 5 种 PSI 蛋白表现出多态性,但在其祖先二倍体物种野生烟草和绒毛状烟草中则没有。这些烟草属植物每个基因组似乎至少有 18 种 PSI 蛋白,分子量范围为 3 至 20 kD。它们包括核基因 psaD、psaE、psaF、psaG、psaH、psaK 和 psaL 的产物、叶绿体基因 psaC 的产物、4.5 kD 和 3.0 kD 的 N 端封闭蛋白,以及一种未鉴定的 12.5 kD 的蛋白。psaD、psaF、psaH 和 psaL 产物各有两种同工型,通过聚丙烯酰胺凝胶电泳中的不同迁移率来区分,psaE 产物有四种同工型。psaD 产物的两种同工型具有不同的氨基酸序列,表明它们由基因组中的不同基因编码。psaE 产物的四种同工型可根据 N 端氨基酸序列分为两组,表明基因组中至少存在两个 psaE 基因。为了研究 PSI 的多态性是否为烟草属植物所特有,我们对番茄(番茄)、拟南芥、红豆(豇豆)和玉米(玉米)的同基因系中的 psaD 和 psaE 产物进行了免疫印迹分析。在番茄、拟南芥和玉米的 psaD 产物中发现了两种电泳上不同的同工型,在番茄、拟南芥和红豆中检测到了 psaE 产物的两种同工型。这些结果表明,除了 psaG 和 psaK 产物外,PSI 的核编码亚基通常有两种同工型。