Date of Award

Fall 1999

Degree Type

Thesis - Restricted

Degree Name

Master of Science (MS)

Department

Chemistry

Abstract

Resonance Raman spectroscopy is now well established as a powerful structural probe of hemeproteins. It can provide detailed structural information for the heme moiety as well as axial ligand disposition. So the interpretation of resonance Raman spectra is very important. The low frequency resonance Raman spectra of P450cam are relatively weak, but rich in the structural information regarding details of the protein-heme interactions. This region has been less systematically studied and definitive assignments require comparison with isotopically labeled samples. So deuteriated heme exchanged at methyl group has been made and was reconstituted with apo-P450cam. Resonance Raman spectra of reconstituted P450cam derivative was studied in order to aid this assignments. Recent attention has focussed on the possible pathway for the proton delivery and possible H-bond donor to stabilize the dioxygen adduct. Since the high reactivity of the ternary dioxygen adduct intermediate make it difficult to have direct study, similar diatomic ligands, such as CO, NO and CN- are used to help to address relative issues. Due to negatively charged, CN- is the best H-bond acceptor and has the advantage to address the H-bond issue. The CN- adducts of reconstituted P450cam were studied and compared with native P450cam CN- adducts. The analysis of the results further confirmed the existence of H-bond donor in the heme active site.

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