Antibodies selected from whole antiserum by fusion proteins as tools for the study of the topology of mitochondrial membrane proteins. Evidence that the N-terminal extremity of the sixth alpha-helix of the uncoupling protein is facing the matrix.
作者： B Miroux , L Casteilla , S Klaus , S Raimbault , S Grandin
DOI: 10.1016/S0021-9258(18)42255-7
关键词:
摘要: The reactivity to freeze-thawed mitochondria or submitochondrial particles of a whole antiserum raised against the uncoupling protein has been investigated. Incubation with brown adipose tissue trapped antibodies reactive toward accessible parts protein. One-third one-half which were present in serum remained free. These highly vesicles obtained by sonication mitochondria, matricial side inner membrane was made accessible. To define epitopes recognized antiserum, different fusion proteins up MalE and fragments used. Immunoaffinity chromatography, using an immobilized purified containing amino acids 253 290 protein, selected specifically directed this part A more precise localization main epitope these is proposed. reacted only particles, indicating orientation epitope. This result, associated other data concerning related mitochondrial carriers such as ADP/ATP translocator phosphate carrier, allowed us determine sixth alpha-helix
sci-hub.st 参考文章(45)
G C Ferreira, R D Pratt, P L Pedersen, Energy-linked anion transport Journal of Biological Chemistry. ,vol. 264, pp. 15628- 15633 ,(1989) , 10.1016/S0021-9258(19)84878-0
H. Aquila, T.A. Link, M. Klingenberg, The uncoupling protein from brown fat mitochondria is related to the mitochondrial ADP/ATP carrier. Analysis of sequence homologies and of folding of the protein in the membrane. The EMBO Journal. ,vol. 4, pp. 2369- 2376 ,(1985) , 10.1002/J.1460-2075.1985.TB03941.X
M. J. Runswick, S. J. Powell, P. Nyren, J. E. Walker, Sequence of the bovine mitochondrial phosphate carrier protein: structural relationship to ADP/ATP translocase and the brown fat mitochondria uncoupling protein. The EMBO Journal. ,vol. 6, pp. 1367- 1373 ,(1987) , 10.1002/J.1460-2075.1987.TB02377.X
Pascale Duplay, Sevec Szmelcman, Hugues Bedouelle, Maurice Hofnung, Silent and functional changes in the periplasmic maltose-binding protein of Escherichia coli K12: I. Transport of maltose☆ Journal of Molecular Biology. ,vol. 194, pp. 663- 673 ,(1987) , 10.1016/0022-2836(87)90243-9
K D Garlid, P Jezek, New substrates and competitive inhibitors of the Cl- translocating pathway of the uncoupling protein of brown adipose tissue mitochondria. Journal of Biological Chemistry. ,vol. 265, pp. 19303- 19311 ,(1990) , 10.1016/S0021-9258(17)30658-0
G C Ferreira, R D Pratt, P L Pedersen, Mitochondrial proton/phosphate transporter. An antibody directed against the COOH terminus and proteolytic cleavage experiments provides new insights about its membrane topology. Journal of Biological Chemistry. ,vol. 265, pp. 21202- 21206 ,(1990) , 10.1016/S0021-9258(17)45346-4
Petr JEZEK, Zdenek DRAHOTA, Sulfhydryl groups of the uncoupling protein of brown adipose tissue mitochondria. Distinction between sulfhydryl groups of the H+ channel and the nucleotide binding site. FEBS Journal. ,vol. 183, pp. 89- 95 ,(1989) , 10.1111/J.1432-1033.1989.TB14900.X
F Bouillaud, J Weissenbach, D Ricquier, Complete cDNA-derived amino acid sequence of rat brown fat uncoupling protein. Journal of Biological Chemistry. ,vol. 261, pp. 1487- 1490 ,(1986) , 10.1016/S0021-9258(17)35962-8
D.L. Murdza-Inglis, H.V. Patel, K.B. Freeman, P. Jezek, D.E. Orosz, K.D. Garlid, Functional reconstitution of rat uncoupling protein following its high level expression in yeast. Journal of Biological Chemistry. ,vol. 266, pp. 11871- 11875 ,(1991) , 10.1016/S0021-9258(18)99038-1
P Duplay, H Bedouelle, A Fowler, I Zabin, W Saurin, M Hofnung, Sequences of the malE gene and of its product, the maltose-binding protein of Escherichia coli K12. Journal of Biological Chemistry. ,vol. 259, pp. 10606- 10613 ,(1984) , 10.1016/S0021-9258(18)91005-7