Exploring the cytochrome c folding mechanism: cytochrome c552 from thermus thermophilus folds through an on-pathway intermediate.
Understanding the role of partially folded intermediate states in the folding mechanism of a protein is a crucial yet very difficult problem. We exploited a kinetic approach to demonstrate that a transient intermediate of a thermostable member of the widely studied cytochrome c family (cytochrome c552 from Thermus thermophilus) is indeed on-pathway. This is the first clear indication of an obligatory intermediate in the folding mechanism of a cytochrome c. The fluorescence properties of this intermediate demonstrate that the relative position of the heme and of the only tryptophan residue cannot correspond to their native orientation. Based on an analysis of the three-dimensional structure of cytochrome c552, we propose an interpretation of the data which explains the residual fluorescence of the intermediate and is consistent with the established role played by some conserved interhelical interactions in the folding of other members of this family. A limited set of topologically conserved contacts may guide the folding of evolutionary distant cytochromes c through the same partially structured state, which, however, can play different kinetic roles, acting either as an intermediate or a transition state.
Travaglini-Allocatelli
Carlo
C
Istituto Pasteur-Fondazione Cenci Bolognetti e Istituto di Biologia e Patologia Molecolari del Consiglio Nazionale delle Ricerche, Dipartimento di Scienze Biochimiche "A. Rossi Fanelli," Università di Roma "La Sapienza," 00185 Rome, Italy.
Gianni
Stefano
S
Morea
Veronica
V
Tramontano
Anna
A
Soulimane
Tewfik
T
Brunori
Maurizio
M
D003574:Cytochrome c Group / Q000737:chemistry*; D045304:Cytochromes c / Q000737:chemistry*; D004305:Dose-Response Relationship, Drug; D019791:Guanidine / Q000494:pharmacology; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D008956:Models, Chemical; D008958:Models, Molecular; D011487:Protein Conformation; D011489:Protein Denaturation; D017510:Protein Folding; D013696:Temperature; D013816:Thermodynamics; D016963:Thermus thermophilus / Q000378:metabolism*
D003574:Cytochrome c Group; C083951:cytochrome c553; D045304:Cytochromes c; C022302:cytochrome C-552; D019791:Guanidine