Inhibition of the CaaX proteases Rce1p and Ste24p by peptidyl (acyloxy)methyl ketones

Stephen B. Porter, Emily R. Hildebrandt, Sarah R. Breevoort, David Z. Mokry, Timothy Dore, Walter K. Schmidt

Research output: Contribution to journalArticle

Abstract

The CaaX proteases Rce1p and Ste24p can independently promote a proteolytic step required for the maturation of certain isoprenylated proteins. Although functionally related, Rce1p and Ste24p are unrelated in primary sequence. They have distinct enzymatic properties, which are reflected in part by their distinct inhibitor profiles. Moreover, Rce1p has an undefined catalytic mechanism, whereas Ste24p is an established zinc-dependent metalloprotease. This study demonstrates that both enzymes are inhibited by peptidyl (acyloxy)methyl ketones (AOMKs), making these compounds the first documented dual specificity inhibitors of the CaaX proteases. Further investigation of AOMK-mediated inhibition reveals that varying the peptidyl moiety can significantly alter the inhibitory properties of AOMKs toward Rce1p and Ste24p and that these enzymes display subtle differences in sensitivity to AOMKs. This observation suggests that this compound class could potentially be engineered to be selective for either of the CaaX proteases. We also demonstrate that the reported sensitivity of Rce1p to TPCK is substrate-dependent, which significantly alters the interpretation of certain reports having used TPCK sensitivity for mechanistic classification of Rce1p. Finally, we show that an AOMK inhibits the isoprenylcysteine carboxyl methyltransferase Ste14p. In sum, our observations raise important considerations regarding the specificity of agents targeting enzymes involved in the maturation of isoprenylated proteins, some of which are being developed as anti-cancer therapeutic agents.

Original languageEnglish (US)
Pages (from-to)853-862
Number of pages10
JournalBiochimica et Biophysica Acta - Molecular Cell Research
Volume1773
Issue number6
DOIs
StatePublished - Jun 1 2007

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Ketones
Peptide Hydrolases
Tosylphenylalanyl Chloromethyl Ketone
Enzymes
Metalloproteases
Protease Inhibitors
Zinc
Proteins
Neoplasms
Therapeutics

Keywords

  • (acyloxy)methyl ketone
  • CaaX protein
  • Isoprenylation
  • Post-translational modification
  • Protease
  • Ras

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology

Cite this

Inhibition of the CaaX proteases Rce1p and Ste24p by peptidyl (acyloxy)methyl ketones. / Porter, Stephen B.; Hildebrandt, Emily R.; Breevoort, Sarah R.; Mokry, David Z.; Dore, Timothy; Schmidt, Walter K.

In: Biochimica et Biophysica Acta - Molecular Cell Research, Vol. 1773, No. 6, 01.06.2007, p. 853-862.

Research output: Contribution to journalArticle

Porter, Stephen B. ; Hildebrandt, Emily R. ; Breevoort, Sarah R. ; Mokry, David Z. ; Dore, Timothy ; Schmidt, Walter K. / Inhibition of the CaaX proteases Rce1p and Ste24p by peptidyl (acyloxy)methyl ketones. In: Biochimica et Biophysica Acta - Molecular Cell Research. 2007 ; Vol. 1773, No. 6. pp. 853-862.
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AU - Dore, Timothy

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