Flexible linkers leash the substrate binding domain of SspB to a peptide module that stabilizes delivery complexes with the AAA plus ClpXP protease

被引:76
作者
Wah, DA
Levchenko, I
Rieckhof, GE
Bolon, DN
Baker, TA
Sauer, RT
机构
[1] MIT, Dept Biol, Cambridge, MA 02139 USA
[2] MIT, Howard Hughes Med Inst, Cambridge, MA 02139 USA
关键词
D O I
10.1016/S1097-2765(03)00272-7
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
SspB dimers bind proteins bearing the ssrA-degradation tag and stimulate their degradation by the ClpXP protease. Here, E. coli SspB is shown to contain a dimeric substrate binding domain of 110-120 N-terminal residues, which binds ssrA-tagged substrates but does not stimulate their degradation. The C-terminal 40-50 residues of SspB are unstructured but are required for SspB to form substrate-delivery complexes with ClpXP. A synthetic peptide containing the 10 C-terminal residues of SspB binds ClpX, stimulates its ATPase activity, and prevents SspB-mediated delivery of GFP-ssrA for ClpXP degradation. This tripartite structure-an ssrA-tag binding and dimerization domain, a flexible linker, and a short peptide module that docks with ClpX-allows SspB to deliver tagged substrates to ClpXP without interfering with their denaturation or degradation.
引用
收藏
页码:355 / 363
页数:9
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