C-TERMINAL SPECIFIC PROTEIN-DEGRADATION - ACTIVITY AND SUBSTRATE-SPECIFICITY OF THE TSP PROTEASE

被引:59
作者
KEILER, KC
SILBER, KR
DOWNARD, KM
PAPAYANNOPOULOS, IA
BIEMANN, K
SAUER, RT
机构
[1] MIT,DEPT BIOL,CAMBRIDGE,MA 02139
[2] MIT,DEPT CHEM,CAMBRIDGE,MA 02139
关键词
INTRACELLULAR PROTEOLYSIS; MASS SPECTROMETRY; PERIPLASMIC PROTEASE; SUBSTRATE SPECIFICITY;
D O I
10.1002/pro.5560040808
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 activity of Tsp, a periplasmic endoprotease of Escherichia coli, has been characterized by assaying the cleavage of protein and peptide substrates, determining the cleavage sites in several substrates, and investigating the kinetics of the cleavage reaction. Tsp efficiently cleaves substrates that have apolar residues and a free alpha-carboxylate at the C-terminus. Tsp cleaves its substrates at a discrete number of sites but with rather broad primary sequence specificity. In addition to preferences for residues at the C-terminus and cleavage sites, Tsp displays a preference for substrates that are not stably folded: unstable variants of Arc repressor are better substrates than a hyperstable mutant, and a peptide with little stable structure is cleaved more efficiently than a protein substrate. These data are consistent with a model in which Tsp cleavage of a protein substrate involves binding to the C-terminal tail of the substrate, transient denaturation of the substrate, and then recognition and hydrolysis of specific peptide bonds.
引用
收藏
页码:1507 / 1515
页数:9
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