Synthetic immunogens. The effect of the conformational space on biological and immunological responses to dimeric hormone constructs.
Chimeras of the double chain bis-cystinyl hinge fragment 225-232/225'-232' of the human IgG1 and of peptides related to human little-gastrin were synthesized, whereby the fully bioactive gastrin sequences 2-17 and 5-17 were amide-bond-linked N- and N- or C-terminally, respectively, to the hinge peptide. All the dimeric constructs proved to be efficient immunogens; however, both the configuration of the constructs and the length of the haptenic gastrin molecule were found to drastically affect the specificity of the antibody response and, thus, the type of dominant immune epitope expressed. The different degree of accessibility of the gastrin chains in the dimers is similarly reflected by their binding affinities to gastrin receptors and their bioactivities in vivo. Molecular dynamics simulations of the chimeric compounds clearly revealed that the conformational space of the gastrin peptide chains 2-17 and 5-17 is strongly restricted upon linkage to the hinge peptide. Only in the gastrin-(2-17) construct does sufficient free conformational space seem to be retained, at least for one of the two gastrin chains, in order to allow folding into the bioactive structure. This also agrees with the observation that the dimeric gastrin-(2-17) behaves like a gastrin monomer in terms of receptor binding affinity and biopotency in vivo; but it could additionally explain why an antibody response of gastrin receptor-like specificity could only be induced with this construct. The experimental data may therefore suggest a high degree of parallelism between the mechanism of recognition of the gastrin peptides in the dimeric constructs as hormonal ligands by the gastrin receptors and as haptens by the immune competent cells.
Moroder
L
L
Max Planck Institut für Biochemie, Martinsried, Federal Republic of Germany.
Musiol
H J
HJ
Köcher
K
K
Bali
J P
JP
Schneider
C H
CH
Guba
W
W
Müller
G
G
Mierke
D F
DF
Kessler
H
H
D000595:Amino Acid Sequence; D000818:Animals; D000917:Antibody Formation; D005755:Gastrins / Q000737:chemistry / Q000276:immunology*; D006168:Guinea Pigs; D006241:Haptens; D007106:Immune Sera / Q000276:immunology*; D007114:Immunization; D009682:Magnetic Resonance Spectroscopy; D008969:Molecular Sequence Data; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D013329:Structure-Activity Relationship
D005755:Gastrins; D006241:Haptens; D007106:Immune Sera