Siggaard-Andersen M, Kauppinen S, von Wettstein-Knowles P
Department of Physiology, Carlsberg Laboratory, Copenhagen, Denmark.
Proc Natl Acad Sci U S A. 1991 May 15;88(10):4114-8. doi: 10.1073/pnas.88.10.4114.
The radioactively labeled beta-ketoacyl thioester synthase inhibitor [3H] cerulenin was used to tag three dimeric barely chloroplast proteins (alpha alpha, alpha beta, and beta beta) from the stromal fraction. Oligonucleotides corresponding to amino acid sequences obtained from the purified proteins were used to generate with the polymerase chain reaction a probe for cDNAs encoding the beta subunit. cDNA sequencing revealed an open reading frame for 462 residues comprising the mature protein and a 35-amino acid transit peptide. The deduced amino acid sequence of the mature protein is homologous to the beta-ketoacyl-[acyl carrier protein] (ACP) synthase I [3-oxoacyl-ACP synthase; acyl-ACP:malonyl-ACP C-acyltransferase (decarboxylating), EC 2.3.1.41] of Escherichia coli. Under analogous experimental conditions [3H]cerulenin tagged a single dimeric protein from spinach chloroplasts.
放射性标记的β-酮脂酰硫酯合酶抑制剂[3H]浅蓝菌素被用于标记来自基质部分的三种二聚体大麦叶绿体蛋白（αα、αβ和ββ）。对应于从纯化蛋白获得的氨基酸序列的寡核苷酸被用于通过聚合酶链反应生成用于编码β亚基的cDNA的探针。cDNA测序揭示了一个由462个残基组成的开放阅读框，包括成熟蛋白和一个35个氨基酸的转运肽。成熟蛋白的推导氨基酸序列与大肠杆菌的β-酮脂酰-[酰基载体蛋白]（ACP）合酶I[3-氧代酰基-ACP合酶；酰基-ACP：丙二酸单酰-ACP C-酰基转移酶（脱羧），EC 2.3.1.41]同源。在类似的实验条件下，[3H]浅蓝菌素标记了来自菠菜叶绿体的一种单一二聚体蛋白。
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