Molecular Genetics and Genomics

, Volume 275, Issue 5, pp 431–436

Substrate specificity of inner membrane peptidase in yeast mitochondria

Original Paper

Abstract

The inner membrane protease (IMP) cleaves intra-organelle sorting peptides from precursor proteins in mitochondria of the yeast Saccharomyces cerevisiae. An unusual feature of the IMP is the presence of two catalytic subunits, Imp1p and Imp2p, which recognize distinct substrate sets even though both enzymes belong to the same protease family. This nonoverlapping substrate specificity was hypothesized to result from the recognition of distinct residues at the P′1 position (also termed +1 position) in the protease substrates. Here, we constructed an extensive series of mutations to obtain a profile of the critical cleavage site residues in IMP substrates and conclude that Imp1p, and not Imp2p, recognizes specific P′1 residues. In addition to its specificity for P′1 residues, Imp1p also shows substrate specificity for the P3 (−3) position. In contrast, Imp2p recognizes the P1 (−1) position and the P3 position. Based on this new understanding of IMP substrate specificity, we conducted a survey for candidate IMP substrates in mammalian mitochondria and found consensus Imp2p cleavage sites in mammalian precursors to cytochrome c1 and glycerol-3-phosphate (G-3-P) dehydrogenase. Presence of a putative Imp2p cleavage site in G-3-P dehydrogenase was surprising, as its yeast ortholog contains an Imp1p cleavage site. To address this issue experimentally, we performed the first co-expression of mammalian IMP with proposed mammalian IMP precursors in yeast and show that murine precursors to cytochrome c1 and G-3-P dehydrogenase are cleaved by murine Imp2p. These results suggest, surprisingly, G-3-P dehydrogenase has switched from Imp1p in yeast to Imp2p in mammals.

Keywords

Inner membrane protease Yeast Signal peptidase Substrate specificity Mitochondria IMMP2L 

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Copyright information

© Springer-Verlag 2006

Authors and Affiliations

  1. 1.Department of Microbiology & ImmunologyVanderbilt University Medical CenterNashvilleUSA

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