C‐terminal specific protein degradation

Activity and substrate specificity of the Tsp protease

Kenneth Charles Keiler, Karen R. Silber, Robert T. Sauer, Kevin M. Downard, Ioannis A. Papayannopoulos, Klaus Biemann

Research output: Contribution to journalArticle

46 Citations (Scopus)

Abstract

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 α‐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.

Original languageEnglish (US)
Pages (from-to)1507-1515
Number of pages9
JournalProtein Science
Volume4
Issue number8
DOIs
StatePublished - Jan 1 1995

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Substrate Specificity
Proteolysis
Degradation
Peptides
Substrates
Proteins
Hydrolysis
Escherichia coli
C-terminal processing peptidase
Denaturation
Kinetics

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology

Cite this

Keiler, K. C., Silber, K. R., Sauer, R. T., Downard, K. M., Papayannopoulos, I. A., & Biemann, K. (1995). C‐terminal specific protein degradation: Activity and substrate specificity of the Tsp protease. Protein Science, 4(8), 1507-1515. https://doi.org/10.1002/pro.5560040808
Keiler, Kenneth Charles ; Silber, Karen R. ; Sauer, Robert T. ; Downard, Kevin M. ; Papayannopoulos, Ioannis A. ; Biemann, Klaus. / C‐terminal specific protein degradation : Activity and substrate specificity of the Tsp protease. In: Protein Science. 1995 ; Vol. 4, No. 8. pp. 1507-1515.
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Keiler, KC, Silber, KR, Sauer, RT, Downard, KM, Papayannopoulos, IA & Biemann, K 1995, 'C‐terminal specific protein degradation: Activity and substrate specificity of the Tsp protease', Protein Science, vol. 4, no. 8, pp. 1507-1515. https://doi.org/10.1002/pro.5560040808

C‐terminal specific protein degradation : Activity and substrate specificity of the Tsp protease. / Keiler, Kenneth Charles; Silber, Karen R.; Sauer, Robert T.; Downard, Kevin M.; Papayannopoulos, Ioannis A.; Biemann, Klaus.

In: Protein Science, Vol. 4, No. 8, 01.01.1995, p. 1507-1515.

Research output: Contribution to journalArticle

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