Antigenic determinants of acylphosphatase from porcine skeletal muscle.
Analysis of the quantitative precipitin reaction of acylphosphatase from porcine skeletal muscle with rabbit antiserum indicated the presence of at least two antigenic determinants on the porcine enzyme molecule. Immunological cross-reactivities of acylphosphatases from equine and rabbit skeletal muscles were examined. In double immunodiffusion with the antiserum, the precipitin lines of the porcine and equine enzymes completely fused, while the rabbit enzyme gave no precipitin line. The reaction between the 125I-labeled porcine enzyme and its antibody was inhibited to the same extent by the porcine and equine enzymes, but not by the rabbit enzyme. The three enzymes were similar in net charge and molecular weight on polyacrylamide gel electrophoreses. No conformational difference among the three enzymes was observed in their circular dichroism spectra. The amino acid composition of the rabbit enzyme differed from those of the porcine and equine enzymes in the contents of Glu, Gly, Lys, and Arg. Differences in the sequence of the rabbit enzyme from that of the porcine enzyme were investigated by comparison of the peptide maps of the tryptic peptides of the two enzymes. Four peptides of the rabbit enzyme were located at different positions from those of the porcine enzyme. Three of the four peptides from both enzymes were sequenced and all the tryptic peptides of both enzymes were characterized by amino acid analysis. The tryptic peptides of rabbit enzyme were tentatively aligned on the basis of their amino acid compositions and sequence homologies, compared with the corresponding peptides of the porcine enzyme. Among five amino acid residues of the porcine enzyme, Arg-4, Asp-28, Arg-31, Glu-56, and Ile-68, which are replaced in the rabbit enzyme, Arg-4 and Asp-28 are considered to be included in the antigenic determinants.
Kizaki
T
T
Mizuno
Y
Y
Takasawa
T
T
Shiokawa
H
H
D017766:Acid Anhydride Hydrolases; D000595:Amino Acid Sequence; D000818:Animals; D002942:Circular Dichroism; D003429:Cross Reactions; D000939:Epitopes; D006736:Horses; D008970:Molecular Weight; D010446:Peptide Fragments / Q000032:analysis; D010744:Phosphoric Monoester Hydrolases / Q000276:immunology*; D011487:Protein Conformation; D011817:Rabbits; D012995:Solubility; D013329:Structure-Activity Relationship; D013552:Swine; D000097024:Acylphosphatase
D017766:Acid Anhydride Hydrolases; D000939:Epitopes; D010446:Peptide Fragments; D010744:Phosphoric Monoester Hydrolases; D000097024:Acylphosphatase