The use of resonance Raman spectroscopy to monitor catalytically important bonds during enzymic catalysis. Application to the hydrolysis of methyl thionohippurate by papain.
Resonance Raman spectra were obtained of the dithioacyl-enzyme intermediate produced during the papain-catalayzed hydrolysis of methyl thionohippurate. Intense resonance Raman features were observed in (formula; see text) and (formula; see text) stretching regions from the intermediate's (formula; see text) chromophore. These results demonstrate that by using a single atom replacement, i.e. sulfur for oxygen, the catalytically crucial bonds in the ester moiety can be monitored during enzymolysis via the resonance Raman spectrum. The method can be extended to other enzymes whose catalytic mechanisms involve the formation of a thiol-acyl intermediate.
D006626:Hippurates; D006868:Hydrolysis; D007700:Kinetics; D010206:Papain / Q000378:metabolism*; D011485:Protein Binding; D013056:Spectrophotometry, Ultraviolet; D013059:Spectrum Analysis, Raman
D006626:Hippurates; C019490:methyl thionohippurate; D010206:Papain