Date of Award
Thesis - Restricted
Master of Science (MS)
Haworth, Daniel T.
Resonance Raman has been used to study the active site of many heme proteins since it allows for selective enhancement of vibrational modes of the chromophoric hemeprosthetic group. The vibrational modes of molecules are quite sensitive to chemical and structural changes; thus structural changes can be monitored by resonance Raman. To date, isotopic labeling has helped to give a definitive set of vibrational modes for the heme prosthetic groups in many heme proteins. Employing reconstituted P450cam derivatives that have been isotopically labeled helps to make assignments of heme modes in the low frequency region, which is relatively weak but rich in structural information. So heme that has its methyl groups deuterated was reconstituted into P450cam and this aided the assignment of some important bands in the low frequency region to propionates. There is a need for more isotopically labeled compounds to make more definitive mode assignments. Isopropyl esters were synthesized, separated and characterized which could be used to make heme with deuterated propionates. Resonance Raman spectroscopy has become an increasingly popular technique for detection of protein intermediates in reaction cycles at cryogenic temperatures using glycerol to trap reactive intermediates of heme proteins. There has been a problem of fluorescence that was obscuring Raman bands in cytochrome P450cam which is due to an impurity in glycerol. Studies were done to purify glycerol.
Manyumwa, Munyaradzi Edith, "Resonance Raman Spectroscopy of Isotopically Labeled Cytochrome P450cam and Low Temperature Measurements" (2000). Master's Theses (1922-2009) Access restricted to Marquette Campus. 2621.