Masking the cryptodeterminant on the 54-kilodalton mouse sperm surface antigen.
Previously, we reported that the epitope of a 54-kDa sperm surface sialoglycoprotein on the flagellum is masked by sialic acid residues. The epitope is referred to as a hidden determinant or cryptodeterminant. This paper reports the manner in which the epitope is masked as evaluated qualitatively and quantitatively by means of SDS-PAGE/immunoblots (Western blot) and ELISA. Immunoblotting with four specific monoclonal antibodies to the 54-kDa sialoglycoprotein--T21 (IgM), MC71 (IgG1), MC81 (IgM), and MC91 (IgM)--demonstrated that not only IgM but also IgG antibody MC71 and the Fab fragment MC71 are masked. Quantitative evaluation with ELISA to compare the antibody titration curves of the masked and unmasked antigens on sialidase-treated and untreated sperm, respectively, indicated that sialidase caused the antibody-binding ability of the epitopes to increase to a different level for each antibody. There were 32-256-, 8-16-, 16-, and 2-4-fold increases in binding to T21, MC71, MC81, and MC91 antibodies, respectively. These results suggest that the antigen-masking through the cryptodeterminant does not depend upon the subtype or the molecular mass of the antibody, but upon the biochemical nature of the epitope region that is closely related to the sialic acid. The mechanism and physiological roles of the antigen-masking are discussed.
Toshimori
K
K
Department of Anatomy, Miyazaki Medical College, Japan.
Araki
S
S
Tanii
I
I
Oura
C
C
D000818:Animals; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000187:drug effects / Q000276:immunology*; D007140:Immunoglobulin Fab Fragments / Q000096:biosynthesis; D007074:Immunoglobulin G / Q000096:biosynthesis; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008813:Mice, Inbred ICR; D008934:Mitogens / Q000494:pharmacology; D009439:Neuraminidase / Q000494:pharmacology; D010504:Periodic Acid / Q000494:pharmacology; D012795:Sialoglycoproteins / Q000276:immunology*; D013094:Spermatozoa / Q000276:immunology*
D000939:Epitopes; D007140:Immunoglobulin Fab Fragments; D007074:Immunoglobulin G; D008934:Mitogens; D012795:Sialoglycoproteins; D010504:Periodic Acid; C009288:metaperiodate; D009439:Neuraminidase