title: Characterization of the tryptophan environments of interleukins 1 alpha and 1 beta by fluorescence quenching and lifetime measurements.
abstract: The tryptophan environments of interleukins 1 alpha and 1 beta, immunomodulatory proteins with similar biological activities but only 25% sequence homology, were characterized by steady-state and dynamic fluorescence measurements. Both proteins exhibited similar emission maxima, but the emission intensity of IL-1 beta was greatly enhanced by increasing the ionic strength of the medium, whereas that of IL-1 alpha was unaffected. The two cytokines were also similarly quenched by the polar quencher acrylamide, but differences were observed for the ionic quenchers iodide and cesium. The fluorescence intensity decays of both cytokines were characterized by two (long and short) component lifetimes. However, the average lifetime of IL-1 beta (4.4 ns) was much longer than that of IL-1 alpha (1.93 ns). Taken together with the results of steady-state measurements, we suggest that the single tryptophan of IL-1 beta is statically quenched by neighboring charged residues, whereas the tryptophan fluorescence of IL-1 alpha is unaffected by ionic strength, and that the tryptophans of the two proteins have different accessibilities to ionic quenchers. The results are discussed in terms of similarities and differences in the tryptophan environments of the two proteins.
authors.affiliation: Physical and Analytical Chemistry, Upjohn Company, Kalamazoo, Michigan 49001.
authors.forename: D E
authors.initials: DE
authors.lastname: Epps
authors.forename: A W
authors.initials: AW
authors.lastname: Yem
authors.forename: M R
authors.initials: MR
authors.lastname: Deibel
mesh_terms: D020106:Acrylamide; D000178:Acrylamides; D002586:Cesium; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007375:Interleukin-1; D007454:Iodides; D007700:Kinetics; D011487:Protein Conformation; D011994:Recombinant Proteins; D013050:Spectrometry, Fluorescence; D014364:Tryptophan
chemical_list: D000178:Acrylamides; D007375:Interleukin-1; D007454:Iodides; D011994:Recombinant Proteins; D002586:Cesium; D020106:Acrylamide; D014364:Tryptophan