Identification of an unusual intein in chloroplast ClpP protease of Chlamydomonas eugametos

被引:36
作者
Wang, SL [1 ]
Liu, XQ [1 ]
机构
[1] DALHOUSIE UNIV,DEPT BIOCHEM,HALIFAX,NS B3H 4H7,CANADA
关键词
D O I
10.1074/jbc.272.18.11869
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 proteasome-like ClpP protease is widely distributed and structurally conserved among bacteria and eukaryotic cell organelles. In Chlamydomonas eugametos, however, the chloroplast clpP gene predicted a much larger ClpP protein containing large insertion sequences (ISs). One insertion sequence, IS2, is 456 amino acid residues long and not similar to known proteins. Here we show that IS2 is an unusual intein, and its protein splicing activity in Escherichia coli cells can be activated by a single amino acid substitution. Analysis of IS2 sequence revealed short sequence motifs that are similar to known intein motifs, including putative LA-GLI-DADG endonuclease motifs. But a histidine residue conserved at the C terminus of known inteins is replaced in the IS2 sequence by a glycine residue (Gly(455)), rendering the IS2 sequence incapable of detectable protein splicing when tested in E. coli cells. Changing Gly(455) to histidine activated the ability of IS2 to undergo protein splicing in E. coli cells. The IS2 sequence (intein) was precisely excised from a precursor protein, with the flanking sequences (exteins) joined together by a normal peptide bond.
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收藏
页码:11869 / 11873
页数:5
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