Optical biosensors study and molecular modeling of interactions between cytochrome P450cam and cytochrome b5

   


1. Institute of Biomedical Chemistry, Russian Academy of Medical Sciences
2. Institut de Biologie Physico Chimique
Type: Experimental/clinical study
UDK:       PubMed Id: 16342666
Year: 2005 vol: 51  issue:5  pages: 501-512
Abstract: The formation of complexes of cytochrome (P450cam) with full-length cytochrome b5 (d-b5) and its trypic water-soluble fragment (t-b5) was analyzed using a two-channel Iasys+ optical biosensor. It was found that t-b5 can form complexes with P450cam, while d-b5 does not interact with P450cam. The invilvement of amine groups of P450cam in the complex formation was demonstrated. The temperature dependence of t-b5im\P450cam complex formation was measured. The association rate constant (kon) increased with temperature, while the dissociation rate constant (koff) practically remained unchanged. It was concluded that hydrophobic interactions play a key role in the complex formation, while electrostatic interactions are significant for complex stabilization. Based on temperature dependence the activation energy, enthalpy and entropy of complex formation were calculated. It was shown that the entropy component plays a key role in t-b5im\P450cam interactions. Computer modeling of P450cam\t-b5 and P450cam\d-b5 interactions was carried out. Using the method of molecular docking some hypotheses of protein-protein complexes were advanced and the best ones were selected based on geometric complementarity, calculated binding energy and probability of electron tunneling between proteins. The computer modeling has shown that only P450cam and t-b5 can form the stable complex. These results are in good agreement with the experimental data obtained with the optical biosensor.
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Reference: Rachenkova N.I., Ivanov Yu.D., Skvortsov V.S., Ivanov A.S., Molnar A.A., Hui Bon Hoa G., Archakov A.I., Optical biosensors study and molecular modeling of interactions between cytochrome P450cam and cytochrome b5, Biomeditsinskaya khimiya, 2005, vol: 51(5), 501-512.
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