Molecular cloning of mouse acid beta-galactosidase cDNA: sequence, expression of catalytic activity and comparison with the human enzyme.
Molecular cloning of mouse acid beta-galactosidase cDNA: sequence, expression of catalytic activity and comparison with the human enzyme.
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DOI：
10.1016/s0006-291x(05)81033-2
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发表时间：
1990-11
影响因子：
3.1
通讯作者：
E. Nanba;K. Suzuki
中科院分区：
文献类型：
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作者：
E. Nanba;K. Suzuki
基于与人基因的同源性，分离了编码小鼠溶酶体酸性β-半乳糖苷酶的全长cDNA。在COS-1细胞表达系统中对4-甲基伞形酮基β-D-半乳糖苷的催化活性为其真实性提供了有力的证据。序列分析表明，人类和小鼠的酶之间的相似性程度约为70%的核苷酸序列和近80%的氨基酸序列。推导的一级氨基酸序列的酶从两个物种表明，在7个可能的N-糖基化位点在人类酶，5个是保守的小鼠酶。在小鼠酶的一级氨基酸序列中发现了三个在人酶中不存在的额外的可能的N-糖基化位点。小鼠酶中的所有七个半胱氨酸残基在人类酶中是保守的。虽然核苷酸序列与E. coli β-半乳糖苷酶的氨基酸序列相似性极小。
A full-length cDNA coding for mouse lysosomal acid β-galatosidase has been isolated on the basis of homology with the human gene. Catalytic activity toward 4-methylumbelliferyl β-D-galactoside in the COS-1 cell expression system provided positive proof for its authenticity. The sequence analysis showed that the degree of similarity between the human and mouse enzymes was approximately 70% in the nucleotide sequence and nearly 80% in the amino acid sequence. The deduced primary amino acid sequences of the enzymes from the two species indicated that, of the seven possible N-glycosylation sites in the human enzyme, five are conserved in the mouse enzyme. Three additional possible N-glycosylation sites, not present in the human enzyme, are found in the primary amino acid sequence of the mouse enzyme. All seven cysteine residues in the mouse enzyme are conserved in the human enzyme. Although the nucleotide sequence could be aligned to 60% identity with the E. coli β-galactosidase, similarity in the amino acid sequence was minimal.