Identification of genes encoding photoconvertible (Class I) water-soluble chlorophyll-binding proteins from Chenopodium ficifolium

被引:1
|
作者
Takahashi, Shigekazu [1 ]
Abe, Eriko [1 ]
Nakayama, Katsumi [1 ]
Satoh, Hiroyuki [1 ]
机构
[1] Toho Univ, Fac Sci, Dept Biomol Sci, Funabashi, Chiba 274, Japan
关键词
Chenopodium ficifolium; chlorophyll; water-soluble chlorophyll-binding protein; Class I WSCP; DUF538; ENERGY-LEVEL STRUCTURE; MOLECULAR-CLONING; INTRACELLULAR-LOCALIZATION; FUNCTIONAL EXPRESSION; ALBUM; PIGMENT; CAULIFLOWER; BRASSICA; COMPLEX; WSCP;
D O I
10.1080/09168451.2014.972326
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Photoconvertible water-soluble chlorophyll-binding proteins, called Class I WSCPs, have been detected in Chenopodiaceae, Amaranthaceae and Polygonaceae plant species. To date, Chenopodium album WSCP (CaWSCP) is the only cloned gene encoding a Class I WSCP. In this study, we identified two cDNAs encoding Chenopodium ficifolium Class I WSCPs, CfWSCP1, and CfWSCP2. Sequence analyses revealed that the open reading frames of CfWSCP1 and CfWSCP2 were 585 and 588bp, respectively. Furthermore, both CfWSCPs contain cystein2 and cystein30, which are essential for the chlorophyll-binding ability of CaWSCP. Recombinant CfWSCP1 and CfWSCP2, expressed in Escherichia coli as hexa-histidine fusion proteins (CfWSCP1-His and CfWSCP2-His), formed inclusion bodies; however, we were able to solubilize these using a buffer containing 8M urea and then refold them by dialysis. The refolded CfWSCP1-His and CfWSCP2-His could bind chlorophylls and exhibited photoconvertibility, confirming that the cloned CfWSCPs are further examples of Class I WSCPs.
引用
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页码:205 / 210
页数:6
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