HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Supplemental Data All Versions of this Article: M600099-MCP200v1 5/8/1426    most recent Submit a response Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Permissions Glossary Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Önder, O. Articles by Daldal, F. Search for Related Content PubMed PubMed Citation Articles by Önder, O. Articles by Daldal, F. Social Bookmarking                      What's this?

Molecular & Cellular Proteomics 5:1426-1436, 2006.
© 2006 by The American Society for Biochemistry and Molecular Biology, Inc.

---

Research

Modifications of the Lipoamide-containing Mitochondrial Subproteome in a Yeast Mutant Defective in Cysteine Desulfurase^*,S

Özlem Önder^,, Heeyong Yoon^,¶, Bianca Naumann, Michael Hippler, Andrew Dancis^¶,|| and Fevzi Daldal^,**

From the Department of Biology, Plant Science Institute and ^¶ Division of Hematology-Oncology, Department of Medicine, School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104

Comparison and identification of mitochondrial matrix proteins from wild-type and cysteine desulfurase-defective (nfs1-14, carrying a hypomorphic allele of NFS1) yeast strains, using two-dimensional gel electrophoresis coupled to mass spectrometry analyses, revealed large changes in the amounts of various proteins. Protein spots that were specifically increased in the nfs1-14 mutant included subunits of lipoamide-containing enzyme complexes: Kgd2, Lat1, and Gcv3, subunits of the mitochondrial -ketoglutarate dehydrogenase, pyruvate dehydrogenase, and glycine cleavage system complexes, respectively. Moreover the increased protein spots corresponded to lipoamide-deficient forms in the nfs1-14 mutant. The increased proteins migrated as separate, cathode-shifted spots, consistent with gain of a lysine charge due to lack of lipoamide addition. Lack of lipoylation of these proteins was further validated using an antibody specific for lipoamide-containing proteins. In addition, this antibody revealed a fourth lipoamide-containing protein, probably corresponding to the E2 component of the branched-chain keto acid dehydrogenase complex. Like the lipoamide-containing forms of Kgd2, Lat1, and Gcv3, this protein also showed decreased lipoic acid reactivity in the nfs1-14 mutant. Cysteine desulfurases, such as yeast NFS1, are required for sulfur addition to iron-sulfur clusters and other sulfur-requiring processes. The results demonstrate that Nfs1 protein is required for the proper post-translational modification of the lipoamide-containing mitochondrial subproteome in yeast and pave the road toward a thorough understanding of its precise role in lipoic acid synthesis.

^** To whom correspondence may be addressed. Tel.: 215-898-4394; E-mail: fdaldal{at}sas.upenn.edu

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				Y. Zhang, E. R. Lyver, E. Nakamaru-Ogiso, H. Yoon, B. Amutha, D.-W. Lee, E. Bi, T. Ohnishi, F. Daldal, D. Pain, et al. Dre2, a Conserved Eukaryotic Fe/S Cluster Protein, Functions in Cytosolic Fe/S Protein Biogenesis Mol. Cell. Biol., September 15, 2008; 28(18): 5569 - 5582. [Abstract] [Full Text] [PDF]

				O. Onder, S. Turkarslan, D. Sun, and F. Daldal Overproduction or Absence of the Periplasmic Protease DegP Severely Compromises Bacterial Growth in the Absence of the Dithiol: Disulfide Oxidoreductase DsbA Mol. Cell. Proteomics, May 1, 2008; 7(5): 875 - 890. [Abstract] [Full Text] [PDF]

				C. Gelling, I. W. Dawes, N. Richhardt, R. Lill, and U. Muhlenhoff Mitochondrial Iba57p Is Required for Fe/S Cluster Formation on Aconitase and Activation of Radical SAM Enzymes Mol. Cell. Biol., March 1, 2008; 28(5): 1851 - 1861. [Abstract] [Full Text] [PDF]