A chimeric mitochondrial precursor protein with internal disulfide bridges blocks import of authentic precursors into mitochondria and allows quantitation of import sites.
作者： G Schatz , D Vestweber
关键词:
摘要: Bovine pancreatic trypsin inhibitor (which contains three intramolecular disulfide bridges) was chemically coupled to the COOH terminus of a purified artificial mitochondrial precursor protein. When resulting chimeric presented energized isolated yeast mitochondria, its moiety prevented protein from completely entering organelle; remained stuck across both membranes, with NH2 in matrix and still exposed on surface. The incompletely imported appeared "jam" import sites since it blocked authentic proteins; did not collapse potential inner membrane. Quantification inhibition indicated that each particle between 10(2) 10(3) sites.
rupress.org
jcb.org参考文章(34)
E.C. Hurt, B. Pesold-Hurt, K. Suda, W. Oppliger, G. Schatz, The first twelve amino acids (less than half of the pre-sequence) of an imported mitochondrial protein can direct mouse cytosolic dihydrofolate reductase into the yeast mitochondrial matrix. The EMBO Journal. ,vol. 4, pp. 2061- 2068 ,(1985) , 10.1002/J.1460-2075.1985.TB03892.X
Albert Haid, Mordechai Suissa, Immunochemical identification of membrane proteins after sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Methods in Enzymology. ,vol. 96, pp. 192- 205 ,(1983) , 10.1016/S0076-6879(83)96017-2
D. Vestweber, G. Schatz, Point mutations destabilizing a precursor protein enhance its post-translational import into mitochondria. The EMBO Journal. ,vol. 7, pp. 1147- 1151 ,(1988) , 10.1002/J.1460-2075.1988.TB02924.X
Masataka MORI, Hiroyuki MATSUE, Satoshi MIURA, Masamiti TATIBANA, Takashi HASHIMOTO, Transport of proteins into mitochondrial matrix. Evidence suggesting a common pathway for 3-ketoacyl-CoA thiolase and enzymes having presequences. FEBS Journal. ,vol. 149, pp. 181- 186 ,(1985) , 10.1111/J.1432-1033.1985.TB08909.X
M Suissa, G Schatz, Import of proteins into mitochondria. Translatable mRNAs for imported mitochondrial proteins are present in free as well as mitochondria-bound cytoplasmic polysomes. Journal of Biological Chemistry. ,vol. 257, pp. 13048- 13055 ,(1982) , 10.1016/S0021-9258(18)33620-2
G Daum, P C Böhni, G Schatz, Import of proteins into mitochondria. Cytochrome b2 and cytochrome c peroxidase are located in the intermembrane space of yeast mitochondria. Journal of Biological Chemistry. ,vol. 257, pp. 13028- 13033 ,(1982) , 10.1016/S0021-9258(18)33617-2
Rod E. Kellems, Venita F. Allison, Ronald A. Butow, Cytoplasmic Type 80 S Ribosomes Associated with Yeast Mitochondria II. EVIDENCE FOR THE ASSOCIATION OF CYTOPLASMIC RIBOSOMES WITH THE OUTER MITOCHONDRIAL MEMBRANE IN SITU Journal of Biological Chemistry. ,vol. 249, pp. 3297- 3303 ,(1974) , 10.1016/S0021-9258(19)42672-0
I.Z. Ades, R.A. Butow, The products of mitochondria-bound cytoplasmic polysomes in yeast. Journal of Biological Chemistry. ,vol. 255, pp. 9918- 9924 ,(1980) , 10.1016/S0021-9258(18)43480-1
G C Shore, R S Hodges, K B Freeman, A T Taneja, L L Gillespie, C Argan, A synthetic signal peptide blocks import of precursor proteins destined for the mitochondrial inner membrane or matrix. Journal of Biological Chemistry. ,vol. 260, pp. 16045- 16048 ,(1985) , 10.1016/S0021-9258(17)36195-1
W J Chen, M G Douglas, The role of protein structure in the mitochondrial import pathway. Unfolding of mitochondrially bound precursors is required for membrane translocation. Journal of Biological Chemistry. ,vol. 262, pp. 15605- 15609 ,(1987) , 10.1016/S0021-9258(18)47769-1