title: Interaction of recombinant human epidermal growth factor with phospholipid vesicles. A steady-state and time-resolved fluorescence study of the bis-tryptophan sequence (Trp49-Trp50).
abstract: The interaction of recombinant human epidermal growth factor with small unilamellar phospholipid vesicles was studied by steady-state and time-resolved fluorescence of the bis-tryptophan sequence (Trp49-Trp50). Steady-state anisotropy measurements demonstrate that strong binding occurred with small unilamellar vesicles made up of acidic phospholipids at acidic pH only (pH < or = 4.7). An apparent stoichiometry for 1,2-dimyristoyl-sn-phosphoglycerol of about 12 phospholipid molecules per molecule of human epidermal growth factor was estimated. The binding appears to be more efficient at temperatures above the gel to liquid-crystalline phase transition. The conformation and the environment of the Trp-Trp sequence are not greatly modified after binding, as judged from the invariance of the excited state lifetime distribution and from that of the fast processes affecting the anisotropy decay. This suggests that the Trp-Trp sequence is not embedded within the bilayer, in contrast to the situation in surfactant micelles (Mayo et al. 1987; Kohda and Inigaki 1992).
authors: Centro de Ingenería Genetica y Biotecnología, Habana, Cuba.
authors: I M
authors: IM
authors: Li De La Sierra
authors: M
authors: M
authors: Vincent
authors: G
authors: G
authors: Padron
authors: J
authors: J
authors: Gallay
mesh_terms: D000595:Amino Acid Sequence; D003001:Cloning, Molecular; D004815:Epidermal Growth Factor / Q000737:chemistry* / Q000235:genetics; D005454:Fluorescence Polarization / Q000379:methods; D006801:Humans; D007700:Kinetics; D008081:Liposomes; D010743:Phospholipids / Q000737:chemistry*; D011485:Protein Binding; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry; D012441:Saccharomyces cerevisiae / Q000235:genetics; D013050:Spectrometry, Fluorescence / Q000379:methods; D013997:Time Factors; D014364:Tryptophan
chemical_list: D008081:Liposomes; D010743:Phospholipids; D011994:Recombinant Proteins; D004815:Epidermal Growth Factor; D014364:Tryptophan