Sigrist M, Staehelin L A
Department of Molecular, Cellular and Developmental Biology, University of Colorado, Boulder, CO.
Biochim Biophys Acta. 1992 Jan 16;1098(2):191-200. doi: 10.1016/s0005-2728(05)80336-6.
The amino acid sequences of more than 40 apoproteins of the light-harvesting complex associated with Photosystem II (LHC II) of various plants have been deduced by sequencing their corresponding genes. These highly conserved sequences fall into two major categories, type 1 and type 2, that differ mainly in a small number of domains close to the N-terminus. We have made polyclonal, monospecific antibodies against synthetic peptides corresponding to the most unique sequence domains of the N-terminal regions of type 1 and type 2 LHC II apoproteins, using sequences derived from petunia genes. On Western blots our anti-type 1 and 2 antibodies crossreact with light-harvesting proteins of petunia, tomato, spinach and several other plants. By using a new gel-system based on ammediol (2-amino-2-methyl-1,3-propanediol), we are able to resolve up to eight LHC II apoproteins. On petunia, tomato and spinach blots the anti type 1 antibodies bind to two or more of the higher molecular weight LHC II polypeptides, whereas the anti type 2 antibodies recognize very specifically only one or two of the lower molecular weight LHC-proteins. In all plants studied, the type 1 LHC II apoproteins are more numerous and span a greater size range than the type 2 apoproteins. This is consistent with the smaller number of type 2 LHC II CAB genes that have been discovered to date.
通过对各种植物中与光系统II（LHC II）相关的捕光复合体的40多种载脂蛋白的相应基因进行测序，已推断出它们的氨基酸序列。这些高度保守的序列分为两大类，即1型和2型，它们主要在靠近N端的少数结构域有所不同。我们利用矮牵牛基因的序列，针对1型和2型LHC II载脂蛋白N端区域最独特的序列结构域合成了多肽，并制备了多克隆单特异性抗体。在蛋白质免疫印迹法中，我们的抗1型和2型抗体与矮牵牛、番茄、菠菜及其他几种植物的捕光蛋白发生交叉反应。通过使用一种基于2-氨基-2-甲基-1,3-丙二醇的新型凝胶系统，我们能够分辨出多达八种LHC II载脂蛋白。在矮牵牛、番茄和菠菜的印迹上，抗1型抗体与两种或更多种较高分子量的LHC II多肽结合，而抗2型抗体则非常特异性地只识别一两种较低分子量的LHC蛋白。在所有研究的植物中，1型LHC II载脂蛋白比2型载脂蛋白数量更多，且分子量范围更广。这与迄今发现的2型LHC II CAB基因数量较少是一致的。
