Identification of a mammalian 10-kDa heat shock protein, a mitochondrial chaperonin 10 homologue essential for assisted folding of trimeric ornithine transcarbamoylase in vitro.
We have identified a 10-kDa stress-inducible mitochondrial protein. The protein is synthesized at elevated rates in cultured rat hepatoma cells challenged with heat shock or amino acid analogues and, therefore, designated heat shock protein 10 (Hsp10). Hsp10 was purified to homogeneity from rat liver and found to exhibit a native molecular mass of 65 kDa, as opposed to a monomeric molecular mass of 10,813.4 +/- 0.41 Da. The amino acid sequence of rat Hsp10 disclosed extensive sequence similarity with bacterial chaperonin (Cpn) 10. Rat Hsp10 and Escherichia coli Cpn60 were used to reconstitute functional trimeric rat ornithine transcarbamoylase from a chemically denatured state with high efficiency. This process depended completely upon rat Hsp10 and was abolished in the presence of a nonhydrolyzable ATP analogue. We conclude that Hsp10 is a eukaryotic Cpn10 homologue and, therefore, together with Cpn60 essential for mitochondrial protein biogenesis. The Cpn-mediated protein-folding apparatus, thus, exhibits a high degree of conservation between prokaryotes and mitochondria of higher eukaryotes.
Department of Biochemistry, La Trobe University, Bundoora, Victoria, Australia.
D J
DJ
Hartman
N J
NJ
Hoogenraad
R
R
Condron
P B
PB
Høj
D000595:Amino Acid Sequence; D000818:Animals; D001419:Bacteria / Q000235:genetics; D001426:Bacterial Proteins / Q000235:genetics*; D018835:Chaperonin 10; D004591:Electrophoresis, Polyacrylamide Gel; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D006360:Heat-Shock Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000302:isolation & purification; D008114:Liver Neoplasms, Experimental / Q000378:metabolism; D046911:Macromolecular Substances; D008715:Methionine / Q000378:metabolism; D008928:Mitochondria / Q000378:metabolism*; D008930:Mitochondria, Liver / Q000378:metabolism*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009954:Ornithine Carbamoyltransferase / Q000737:chemistry / Q000378:metabolism*; D011487:Protein Conformation; D011489:Protein Denaturation; D051381:Rats; D012689:Sequence Homology, Nucleic Acid; D014407:Tumor Cells, Cultured
D001426:Bacterial Proteins; D018835:Chaperonin 10; D006360:Heat-Shock Proteins; D046911:Macromolecular Substances; D008715:Methionine; D009954:Ornithine Carbamoyltransferase