Solution structure of the acetylated and noncleavable mitochondrial targeting signal of rat chaperonin 10

J. A. Jarvis, M. T. Ryan, N. J. Hoogenraad, D. J. Craik, P. B. Hoj

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Abstract

Chaperonin 10 (Cpn10) is one of only a few mitochondrial matrix proteins synthesized without a cleavable targeting signal. Using a truncated form of Cpn10 and synthetic peptides in mitochondrial import assays, we show that the N-terminal region is both necessary and sufficient for organellar targeting in vitro. To elucidate the structural features of this topogenic signal, peptides representing residues 1-25 of rat Cpn10 were synthesized with and without the naturally occurring N-terminal acetylation. 1H NMR spectroscopy in 20% CF3CH2OH, H2O showed that both peptides assume a stable helix- turn-helix motif and are highly amphiphilic in nature. Chemical shift and coupling constant data revealed that the N-terminal helix is stabilized by N- acetylation, whereas NOE and exchange studies were used to derive a three dimensional structure for the acetylated peptide. Those findings are discussed with respect to a recent model predicting that targeting sequences forming a continuous α-helix of more than 11 residues cannot adopt a conformation necessary for proteolysis by the matrix located signal peptidases (Hammen, P. K., Gorenstein, D. G., and Weiner, H. (1994) Biochemistry 33, 8610-8617).

Original languageEnglish
Pages (from-to)1323-1331
Number of pages9
JournalThe Journal of Biological Chemistry
Volume270
Issue number3
DOIs
Publication statusPublished - 1 Jan 1995
Externally publishedYes

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