title: Fluorescence studies with human epidermal growth factor.
abstract: Steady-state and time-resolved fluorescence studies have been performed with human epidermal growth factor, a small globular protein having two adjacent tryptophan residues near its C-terminus. Based on the relatively red fluorescence and accessibility to solute quenchers, the two tryptophan residues are found to be exposed to solvent. Anisotropy decay measurements show the dominant depolarizing process to have a sub-nanosecond rotational correlation time indicating the existence of rapid segmental motion of the fluorescing tryptophan residues. From an analysis of the low-temperature excitation anisotropy spectrum of the protein (and in comparison with that of tryptophan, the peptide melittin, and the dipeptide trp-trp), it is concluded that homo-energy transfer and/or exciton interaction occurs between the adjacent tryptophan residues. A thermal transition in the structure of the protein, which is observed by circular dichroism measurements, is not sensed by the steady-state fluorescence of the protein. This result, in conjunction with the anisotropy decay results, indicates that the two tryptophan residues are in a highly flexible C-terminus segment, which is not an integral part of the three-dimensional structure of the protein. Fluorescence measurements with three site-directed mutants also show very little variation.
authors.affiliation: Department of Biochemistry, University of Missouri, Columbia 65211.
authors.forename: C A
authors.initials: CA
authors.lastname: Ghiron
authors.forename: M R
authors.initials: MR
authors.lastname: Eftink
authors.forename: D A
authors.initials: DA
authors.lastname: Engler
authors.forename: S K
authors.initials: SK
authors.lastname: Niyogi
mesh_terms: D000595:Amino Acid Sequence; D004815:Epidermal Growth Factor / Q000737:chemistry* / Q000235:genetics; D006801:Humans; D049449:Luminescence; D016297:Mutagenesis, Site-Directed; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry; D013050:Spectrometry, Fluorescence
chemical_list: D011994:Recombinant Proteins; D004815:Epidermal Growth Factor