Volokita M, Somerville C R
MSU-DOE Plant Research Laboratory, East Lansing 48824.
J Biol Chem. 1987 Nov 25;262(33):15825-8.
A cDNA clone encoding the peroxisomal enzyme glycolate oxidase (EC 1.1.3.15) was identified by probing a cDNA library of spinach with synthetic oligonucleotides based on the partial amino acid sequence of the enzyme. Determination of the DNA sequence of the 1526-nucleotide cDNA indicated a 1107-nucleotide open reading frame which encodes a polypeptide of 40,282 daltons. The polypeptide produced by in vitro transcription and translation of the cDNA insert had the same apparent subunit molecular mass as the enzyme purified from leaves, indicating that the cDNA encodes a full-length polypeptide and that no cleavage of the polypeptide is required for uptake of the polypeptide by peroxisomes. Comparison of the deduced amino acid sequence with those of two other plant peroxisomal proteins revealed a region of homology which may be involved in directing proteins to the peroxisome.
通过用基于该酶部分氨基酸序列的合成寡核苷酸探测菠菜的cDNA文库，鉴定出了一个编码过氧化物酶体酶乙醇酸氧化酶（EC 1.1.3.15）的cDNA克隆。对1526个核苷酸的cDNA的DNA序列测定表明，有一个1107个核苷酸的开放阅读框，其编码一个40282道尔顿的多肽。由cDNA插入片段的体外转录和翻译产生的多肽，其表观亚基分子量与从叶片中纯化的酶相同，这表明该cDNA编码一个全长多肽，并且该多肽被过氧化物酶体摄取不需要进行切割。将推导的氨基酸序列与其他两种植物过氧化物酶体蛋白的序列进行比较，发现了一个可能参与将蛋白质导向过氧化物酶体的同源区域。
