Heterogeneity of hepatic tyrosine aminotransferase. Separation of the multiple forms from rat and frog liver by isoelectric focussing and hydroxylapatite column chromatography and their partial characterization.
L-Tyrosine:2-oxoglutarate aminotransferase (EC 2.6.1.5; TAT) and other enzymes that transaminate tyrosine in rat liver cytosol have been separated into four fractions by isoelectric focussing. One of the forms is probably identical to mitochondrial L-aspartate:2-oxoglutarate aminotransferase (EC 2.6.1.1.; mASAT). The other three forms have pI's of 4.72, 4.98 and 5.30 and Km values of 1.3 and 0.3 mM for tyrosine and alpha-ketoglutarate. These heat stable forms have little or no ASAT activity. Rat liver TAT is also separated into three peaks by hydroxylapatite. Each fraction gives only one peak of activity when electrofocussed separately. In the frog, three groups of peaks of TAT activity have been separated by hydroxylapatite column chromatography. The first group is connected with ASAT activity. These peaks (pI's 6.35, 6.50 and 6.90) are heat stable and have a Km value for tyrosine of 4 mM. These fractions probably represent cytoplasmic ASAT (sASAT). The second group of peaks has at least two subforms (pI's 9.0 and 9.4, Km for tyrosine 15 mM). These forms probably represent mASAT. The third group consists of three forms that resemble the major forms of rat liver TAT. These results indicate that heterogeneity is common to many aminotransferases and independent of regulation by glucocorticoids.
Heterogeneity of hepatic tyrosine aminotransferase. Separation of the multiple forms from rat and frog liver by isoelectric focussing and hydroxylapatite column chromatography and their partial characterization. L-Tyrosine:2-oxoglutarate aminotransferase (EC 2.6.1.5; TAT) and other enzymes that transaminate tyrosine in rat liver cytosol have been separated into four fractions by isoelectric focussing. One of the forms is probably identical to mitochondrial L-aspartate:2-oxoglutarate aminotransferase (EC 2.6.1.1.; mASAT). The other three forms have pI's of 4.72, 4.98 and 5.30 and Km values of 1.3 and 0.3 mM for tyrosine and alpha-ketoglutarate. These heat stable forms have little or no ASAT activity. Rat liver TAT is also separated into three peaks by hydroxylapatite. Each fraction gives only one peak of activity when electrofocussed separately. In the frog, three groups of peaks of TAT activity have been separated by hydroxylapatite column chromatography. The first group is connected with ASAT activity. These peaks (pI's 6.35, 6.50 and 6.90) are heat stable and have a Km value for tyrosine of 4 mM. These fractions probably represent cytoplasmic ASAT (sASAT). The second group of peaks has at least two subforms (pI's 9.0 and 9.4, Km for tyrosine 15 mM). These forms probably represent mASAT. The third group consists of three forms that resemble the major forms of rat liver TAT. These results indicate that heterogeneity is common to many aminotransferases and independent of regulation by glucocorticoids.