Characteristics of aspartate aminotransferase binding immunoglobulin determined by the isotope method.
A woman who had no known underlying diseases showed a persistent elevation (about 300 U/L) of serum aspartate aminotransferase (AST) without other abnormal laboratory findings. Cellolose gel electrophoresis showed that the AST activity in the patient had an atypical band with slower mobility than normal AST. When the sera from the patient and from a patient with acute hepatitis were mixed, the atypical band increased in density and the band of normal size AST disappeared. When the serum was fractionated on Sephadex G-200 gel filtration medium, almost all AST activity was found between the void volume and the gamma-globulin fraction. However, the AST activity in this fraction was not retained on dissociation into small AST by acid treatment. This suggests the loss of enzyme activity in dissociated small AST. The patient's serum was then incubated with iodine 125-labeled porcine AST; when this was fractionated on gel filtration medium, the main radioactivity was eluted in the void volume fraction. The binding activity for 125I-porcine AST was found in the gamma-globulin fraction obtained by gel filtration. The affinity constant of 125I-porcine AST binding to the gamma-globulin fraction was 1.0 x 10(-8) mol/L by Scatchard analysis. The binding gamma-globulin appeared to be (polyclonal) IgG, and the binding site was located in F(ab')2 and Fab fragments. The IgG could be bound with both human and porcine AST but not with chick AST. Thus the IgG appears specific for AST of mammalian species.(ABSTRACT TRUNCATED AT 250 WORDS)
Ohtsuki
T
T
Central Clinical Laboratory, Shiga University of Medical Science, Japan.
Inui
T
T
Andoh
A
A
Chen
W
W
Ochi
Y
Y
Sasaki
M
M
Kimura
T
T
Hosoda
S
S
D000328:Adult; D000818:Animals; D001219:Aspartate Aminotransferases / Q000097:blood / Q000378:metabolism*; D002645:Chickens; D002850:Chromatography, Gel / Q000379:methods*; D004586:Electrophoresis / Q000379:methods*; D005260:Female; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000032:analysis / Q000378:metabolism; D007147:Immunoglobulin Light Chains / Q000032:analysis / Q000378:metabolism; D007136:Immunoglobulins / Q000032:analysis* / Q000378:metabolism*; D007457:Iodine Radioisotopes; D011485:Protein Binding; D013552:Swine
D007140:Immunoglobulin Fab Fragments; D007147:Immunoglobulin Light Chains; D007136:Immunoglobulins; D007457:Iodine Radioisotopes; D001219:Aspartate Aminotransferases