Cloning and expression of mouse legumain, a lysosomal endopeptidase

被引:123
作者
Chen, JM [1 ]
Dando, PM [1 ]
Stevens, RAE [1 ]
Fortunato, M [1 ]
Barrett, AJ [1 ]
机构
[1] Babraham Inst, MRC, Peptidase Lab, Cambridge CB2 4AT, England
关键词
D O I
10.1042/bj3350111
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
Legumain, a recently discovered mammalian cysteine endopeptidase, was found in all mouse tissues examined, but was particularly abundant in kidney and placenta. The distribution in subcellular fractions of mouse and rat kidney showed a lysosomal localization, and activity was detectable only after the organelles were disrupted. Nevertheless, ratios of legumain activity to that of cathepsin B differed considerably between mouse tissues, cDNA encoding mouse legumain was cloned and sequenced, the deduced amino acid sequence proving to be 83 % identical to that of the human protein [Chen, Dando, Rawlings, Brown, Young, Stevens, Hewitt, Watts and Barrett (1997) J. Biol. Chem. 272, 8090-8098]. Recombinant mouse legumain was expressed in human embryonic kidney 293 cells by use of a Vector containing a cytomegalovirus promoter. The recombinant enzyme was partially purified and found to be an asparagine-specific endopeptidase closely similar to naturally occurring pig kidney legumain.
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页码:111 / 117
页数:7
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