Protein fragment complementation in M.HhaI DNA methyltransferase

被引:19
作者
Choe, W
Chandrasegaran, S
Ostermeier, M
机构
[1] Johns Hopkins Univ, Bloomberg Sch Publ Hlth, Dept Environm Hlth Sci, Baltimore, MD 21205 USA
[2] Johns Hopkins Univ, Dept Chem & Biomol Engn, Baltimore, MD 21218 USA
关键词
M.HhaI; DNA methyltransferase; protein fragment complementation; leucine zipper; incremental truncation; protein engineering; M.AquI; M.BssHII; M.BspRI; DNA methylation;
D O I
10.1016/j.bbrc.2005.07.017
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
The 5mC DNA methyltransferase M.HhaI can be split into two individually inactive N- and C-terminal fragments that together can form an active enzyme in vivo capable of efficiently methylating DNA. This active fragment pair was identified by creating libraries of M.HhaI gene fragment pairs and then selecting for the pairs that code for an active 5mC methyltransferase. The site of bisection for successful protein fragment complementation in M.HhaI was in the variable region near the target recognition domain between motif VIII and TRD. This same region is the location of bifurcation in the naturally split 5mC methyltransferase M.AquI, the location for circular permutation in M.BssHII, and the location for previously engineered split versions of M.BspRI. (c) 2005 Elsevier Inc. All rights reserved.
引用
收藏
页码:1233 / 1240
页数:8
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