Pichersky E, Subramaniam R, White M J, Reid J, Aebersold R, Green B R
Biology Department, University of Michigan, Ann Arbor 48109.
Mol Gen Genet. 1991 Jun;227(2):277-84. doi: 10.1007/BF00259681.
CP29, the core chlorophyll a/b (CAB) antenna complex of Photosystem II (PSII), has two nuclear-encoded polypeptides of approximately 26 and 28 kDa in tomato (Lycopersicon esculentum). Cab9, the gene for the Type I (26 kDa) CP29 polypeptide was cloned by immunoscreening a tomato leaf cDNA library. Its identity was confirmed by sequencing tryptic peptides from the mature protein. Cab9 is a single-copy gene with five introns, the highest number found in a CAB protein. In vitro transcription-translation gave a 31 kDa precursor which was cleaved to about 26 kDa after import into isolated tomato chloroplasts. The Cab9 polypeptide has the two highly conserved regions common to all CAB polypeptides, which define the members of this extended gene family. Outside of the conserved regions, it is only slightly more closely related to other PSII CABs than to PSI CABs. Sequence analysis of tryptic peptides from the Type II (28 kDa) CP29 polypeptide showed that it is also a member of the CAB family and is very similar or identical to the CP29 polypeptide previously isolated from spinach. All members of the CAB family have absolutely conserved His, Gln and Asn residues which could ligate the Mg atoms of the chlorophylls, and a number of conserved Asp. Glu, Lys and Arg residues which could form H-bonds to the polar groups on the porphyrin rings. The two conserved regions comprise the first and third predicted trans-membrane helices and the stroma-exposed segments preceding them.
CP29是光系统II(PSII)的核心叶绿素a/b(CAB)天线复合体,在番茄(Lycopersicon esculentum)中有两个核编码的多肽,分子量约为26 kDa和28 kDa。通过免疫筛选番茄叶cDNA文库,克隆了I型(26 kDa)CP29多肽的基因Cab9。通过对成熟蛋白的胰蛋白酶肽段进行测序,证实了其身份。Cab9是一个单拷贝基因,有五个内含子,这是在CAB蛋白中发现的最多数量。体外转录-翻译产生了一个31 kDa的前体,导入分离的番茄叶绿体后被切割成约26 kDa。Cab9多肽具有所有CAB多肽共有的两个高度保守区域,这些区域定义了这个扩展基因家族的成员。在保守区域之外,它与其他PSII CAB的关系仅比与PSI CAB的关系稍近一些。对II型(28 kDa)CP29多肽的胰蛋白酶肽段进行序列分析表明,它也是CAB家族的成员,并且与先前从菠菜中分离的CP29多肽非常相似或相同。CAB家族的所有成员都有绝对保守的His、Gln和Asn残基,它们可以连接叶绿素的Mg原子,还有一些保守的Asp、Glu、Lys和Arg残基,它们可以与卟啉环上的极性基团形成氢键。这两个保守区域包括预测的第一和第三个跨膜螺旋以及它们之前暴露于基质的片段。