Immunodominant structures of human growth hormone identified by homolog-scanning mutagenesis.
Homolog-scanning mutagenesis has been reported to be useful in elucidating the antigenic epitopes recognized by monoclonal antibodies and hGH binding to its receptor. However, little is known about which structures are recognized as immunodominant by murine serum antibodies. Therefore, the previously published series of hGH homologs and additional mutants of human placental lactogen (hPL), porcine growth hormone (pGH), and human prolactin (hPRL) were examined for their interaction with murine serum derived anti-hGH antibodies. As compared to wild-type hGH, nine of the nineteen segment substituted mutants tested showed a significant reduction in binding to anti-hGH sera. These disruptive substitutions mapped to 5 regions on a structural model of hGH: the length of helix 1 (residues 11-33), the loop between the first disulfide bond and helix 2 (residues 54-74), the beginning of helix 3 (residues 109-112), the carboxyl half of helix 4 (residues 167-182), and the final carboxyl terminus segment of the molecule (residues 184-191). In terms of the current structural model, three of the five immunodominant regions (the loop between residues 54-74, central portion of helix 4 to the carboxyl terminus and part of the amino terminus region of helix 1) closely overlaps the hGH receptor binding epitopes.
Weber
J R
JR
Department of Immunobiology, Genentech Inc., South San Francisco, CA 94080.
Nelson
C
C
Cunningham
B C
BC
Wells
J A
JA
Fong
S
S
D000595:Amino Acid Sequence; D000818:Animals; D001666:Binding Sites, Antibody; D001667:Binding, Competitive / Q000276:immunology; D005260:Female; D013006:Growth Hormone / Q000737:chemistry / Q000276:immunology*; D006801:Humans; D016056:Immunodominant Epitopes / Q000235:genetics* / Q000276:immunology; D007074:Immunoglobulin G / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D016296:Mutagenesis; D012689:Sequence Homology, Nucleic Acid
D016056:Immunodominant Epitopes; D007074:Immunoglobulin G; D013006:Growth Hormone