Spectral characterization of environment-sensitive adducts of interleukin-1 beta.
We have determined the fluorescence properties of two covalently attached acrylodan derivatives of recombinant human interleukin-1 beta, namely the Cys-8 and Lys-103 adducts. The emission and excitation maxima indicated the presence of two operationally distinct conformers for each probe. The iodide quenching and the kinetics of fluorescence changes associated with guanidinium chloride-induced denaturation show that each covalent adduct exists both in hydrated and dehydrated environments. Furthermore, fluorescence changes associated with the binding of the adducts to a recombinant soluble murine receptor indicated that only the conformers with the label in the hydrophobic environment are competent toward the soluble murine interleukin receptor and that the hydrated and dehydrated conformers are in a dynamic equilibrium on the time scale of the binding experiments.
Epps
D E
DE
Upjohn Laboratories, Upjohn Company, Kalamazoo, Michigan 49001.
Yem
A W
AW
Fisher
J F
JF
McGee
J E
JE
Paslay
J W
JW
Deibel
M R
MR
D015081:2-Naphthylamine / Q000031:analogs & derivatives* / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D003545:Cysteine; D006801:Humans; D007375:Interleukin-1 / Q000737:chemistry* / Q000378:metabolism; D007700:Kinetics; D008239:Lysine; D051379:Mice; D011487:Protein Conformation; D011971:Receptors, Immunologic / Q000187:drug effects / Q000378:metabolism; D017472:Receptors, Interleukin-1; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D013050:Spectrometry, Fluorescence
D007375:Interleukin-1; D011971:Receptors, Immunologic; D017472:Receptors, Interleukin-1; D011994:Recombinant Proteins; C038518:acrylodan; D015081:2-Naphthylamine; D008239:Lysine; D003545:Cysteine