Immunologically reactive tryptic fragments of human prostatic acid phosphatase.
Three peptide fragments (designated II, III and IV) of human prostatic acid phosphatase (PAP) were isolated to homogeneity from a limited tryptic hydrolysate of PAP by gel filtration on Sephadex G-100, followed by chromatography on DEAE-cellulose and Sephadex G-75. The homogeneity was confirmed by disc poly-acrylamide-gel electrophoresis. The Mr values were 32 500, 25 000 and 11 000 as estimated by gel filtration and sodium dodecyl sulphate/polyacrylamide-gel electrophoresis. Immunoprecipitation study revealed that only fragment II formed an immune precipitate with anti-PAP antibodies. Fragment II exhibited 45% of maximum inhibitory activity on the reaction between PAP and goat anti-PAP IgG (immunoglobulin G) antibodies (or rabbit anti-PAP antibodies), whereas fragments III and IV demonstrated 24% (or 23%) and 29% (or 27%) inhibition respectively. A mixture of these three tryptic fragments of PAP result in 96% (for goat anti-PAP antibodies) and 94% (for rabbit anti-PAP antibodies) inhibitory activities, which were equivalent to the sum of maximum inhibitory activity of the three fragments individually. The results demonstrated that these three tryptic peptide fragments carried all the antigenic active sites of the native PAP, and suggested that the entire molecule of human PAP comprised a minimum of four distinguishable, nonoverlapping antigenic determinants. These three fragments also were shown to retain all the disulphide bonds of the native PAP, and thus were useful reagents for the elucidation of PAP molecular structure.
D000135:Acid Phosphatase / Q000276:immunology*; D000596:Amino Acids / Q000032:analysis; D000906:Antibodies / Q000276:immunology; D002850:Chromatography, Gel; D004591:Electrophoresis, Polyacrylamide Gel; D000939:Epitopes / Q000276:immunology; D006801:Humans; D007122:Immunoelectrophoresis; D007074:Immunoglobulin G / Q000276:immunology; D008297:Male; D010446:Peptide Fragments / Q000276:immunology*; D010455:Peptides / Q000276:immunology*; D011467:Prostate / Q000201:enzymology; D014357:Trypsin
D000596:Amino Acids; D000906:Antibodies; D000939:Epitopes; D007074:Immunoglobulin G; D010446:Peptide Fragments; D010455:Peptides; D000135:Acid Phosphatase; D014357:Trypsin