In Search of Circular Permuted Variants of Escherichia coli Dihydrofolate Reductase
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概要
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A circularized form of a Cys-free mutant of Escherichia coli dihydrofolate reductase (DHFR) was used to search for a proteoltic site that gave new N- and C-termini on circularized DHFR with enzyme activity. Of the six sitespecific proteolytic enzymes tested, three proteases, Achromobacter protease I (lysine-specific endopeptidase), asparaginylendopeptidase, and Staphylococcus aureus V8 protease, cleaved a single site of a the circularized DHFR to form circular permuted variants. Twenty-four possible sites for cleavage were found formation of eight circular permuted variants was suggested by results of N-terminal sequence analysis of the linearized proteins isolated by gel filtration in the presence of 5 M guanidine hydrochloride. Mapping of the predicted cleavage sites on the DHFR molecule suggested that they were not all at a specific loop and, therefore, there are many possible circular permuted variants.
- 社団法人日本農芸化学会の論文
- 1998-04-23
著者
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Iwakura Masahiro
National Institute of Advanced Industrial Science and Technology
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Iwakura Masahiro
National Institute Of Bioscience And Human Technology
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Iwakura Masahiro
National Institute For Advanced Interdisciplinary Research
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