title: Fluorescence lifetime studies with staphylococcal nuclease and its site-directed mutant. Test of the hypothesis that proline isomerism is the basis for nonexponential decays.
abstract: Using frequency domain methods, the fluorescence decay of Trp-140 in staphylococcal nuclease and its site-directed mutant (Pro-117----Gly) has been examined. Based on nuclear magnetic resonance (NMR) studies (Evans, P. A., C. M. Dobson, R. A. Kautz, G. Hatfull, and R. O. Fox. 1987. Nature [Lond.]. 329:266-268), it is believed that nuclease exists in two macroscopic, native conformations and that the slow interconversion of these conformations is controlled by the cis----trans isomerization of Pro-117. The above mutant shows only one native conformation in NMR experiments. To test the hypothesis that the biexponential fluorescence decay of Trp-140 of nuclease can also be related to the existence of these conformational states of the protein, we have compared the decay patterns of the wild type and mutant. Essentially no difference was observed, which indicates that there is some other basis for the nonexponential decay of Trp-140. We have used global nonlinear least squares analysis to link the fit of data at several temperatures.
authors.affiliation: Department of Chemistry, University of Mississippi, University 38677.
authors.forename: M R
authors.initials: MR
authors.lastname: Eftink
authors.forename: C A
authors.initials: CA
authors.lastname: Ghiron
authors.forename: R A
authors.initials: RA
authors.lastname: Kautz
authors.forename: R O
authors.initials: RO
authors.lastname: Fox
mesh_terms: D001665:Binding Sites; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics; D005796:Genes; D005798:Genes, Bacterial; D005998:Glycine; D008836:Micrococcal Nuclease / Q000235:genetics / Q000378:metabolism*; D009154:Mutation; D011392:Proline; D013050:Spectrometry, Fluorescence; D013211:Staphylococcus aureus / Q000201:enzymology / Q000235:genetics
chemical_list: D011392:Proline; D008836:Micrococcal Nuclease; D005998:Glycine