title: Dynamics in the transient complex of plastocyanin-cytochrome f from Prochlorothrix hollandica.
abstract: The nature of transient protein complexes can range from a highly dynamic ensemble of orientations to a single well-defined state. This represents variation in the equilibrium between the encounter and final, functional state. The transient complex between plastocyanin (Pc) and cytochrome f (cytf) of the cyanobacterium Prochlorothrix hollandica was characterized by NMR spectroscopy. Intermolecular pseudocontact shifts and chemical shift perturbations were used as restraints in docking calculations to determine the structure of the wild-type Pc-cytf complex. The orientation of Pc is similar to orientations found in Pc-cytf complexes from other sources. Electrostatics seems to play a modest role in complex formation. A large variability in the ensemble of lowest energy structures indicates a dynamic nature of the complex. Two unusual hydrophobic patch residues in Pc have been mutated to the residues found in other plastocyanins (Y12G/P14L). The binding constants are similar for the complexes of cytf with wild-type Pc and mutant Pc, but the chemical shift perturbations are smaller for the complex with mutant Pc. Docking calculations for the Y12G/P14L Pc-cytf complex did not produce a converged ensemble of structures. Simulations of the dynamics were performed using the observed averaged NMR parameters as input. The results indicate a surprisingly large amplitude of mobility of Y12G/P14L Pc within the complex. It is concluded that the double mutation shifts the complex further from the well-defined toward the encounter state.
authors.affiliation: Leiden Institute of Chemistry, Leiden University, Gorlaeus Laboratories, P.O. Box 9502, 2300 RA Leiden, The Netherlands.
authors.forename: Rinske
authors.initials: R
authors.lastname: Hulsker
authors.forename: Maria V
authors.initials: MV
authors.lastname: Baranova
authors.forename: George S
authors.initials: GS
authors.lastname: Bullerjahn
authors.forename: Marcellus
authors.initials: M
authors.lastname: Ubbink
mesh_terms: D002104:Cadmium / Q000737:chemistry; D003198:Computer Simulation; D003300:Copper / Q000737:chemistry; D045348:Cytochromes f / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D009682:Magnetic Resonance Spectroscopy / Q000379:methods / Q000592:standards; D008956:Models, Chemical; D008968:Molecular Conformation; D010970:Plastocyanin / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D019692:Prochlorothrix / Q000737:chemistry* / Q000201:enzymology*; D012015:Reference Standards
chemical_list: D002104:Cadmium; D003300:Copper; D010970:Plastocyanin; D045348:Cytochromes f