CD and fluorescence studies of the human granulocyte-macrophage colony-stimulating factor and related peptide conformations in aqueous solution.
The absorption, CD, and fluorescence emission spectra, and the fluorescence emission and depolarization lifetimes of the human granulocyte-macrophage colony-stimulating factor (hGM-CSF) and related peptides previously tested for their immunological activity, were measured in water at various pHs and temperatures to obtain information on their conformation in solution. The aim was to correlate the amino acid sequences, and the chain conformations and dynamics of the peptides, with their immunological properties. The CD spectrum of hGM-CSF revealed, as expected, a structure in solution similar to that in the crystalline state, but the fluorescence data suggest that the Trp 122 residue is more accessible to the solvent than the x-ray data would lead one to expect. They also suggest that some flexibility exists between the protein's two domains, one made up of the alpha-helices A and C and the other of the alpha-helices B and D plus the two beta-strands. In aqueous solution, none of the tested peptide CD spectra could be linked to a recognizable ordered conformation, i.e., an alpha-helix or a beta-sheet. The fluorescence of the peptide 11-24 suggests that the Trp 13 residue may appear in two types of situations: (a) in aqueous solution and (b) within a globular structure. Its CD spectra show that the tryptophan residue exists in both cases in a highly asymmetric environment independent of the pH.
Casté
L
L
Dipartimento di Chimica e Chimica Industriale, Università di Pisa, Italy.
Zandomeneghi
M
M
Houben
J L
JL
Rovero
P
P
Pegoraro
S
S
Revoltella
P R
PR
Brochon
J C
JC
D002942:Circular Dichroism; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000032:analysis*; D006801:Humans; D010455:Peptides / Q000032:analysis*; D011487:Protein Conformation; D012996:Solutions; D013050:Spectrometry, Fluorescence; D014867:Water
D010455:Peptides; D012996:Solutions; D014867:Water; D016178:Granulocyte-Macrophage Colony-Stimulating Factor