title: Identification of mitochondrial branched chain aminotransferase and its isoforms in rat tissues.
abstract: The tissue distribution and subcellular location of branched chain aminotransferase was analyzed using polyclonal antibodies against the enzyme purified from rat heart mitochondria (BCATm). Immunoreactive proteins were visualized by immunoblotting. The antiserum recognized a 41-kDa protein in the 100,000 x g supernatant from a rat heart mitochondrial sonicate. The 41-kDa protein was always present in mitochondria which contained branched chain aminotransferase activity, skeletal muscle, kidney, stomach, and brain, but not in cytosolic fractions. In liver mitochondria, which have very low levels of branched chain aminotransferase activity, the 41-kDa protein was not present. However, two immunoreactive proteins of slightly higher molecular masses were identified. These proteins were located in hepatocytes. The 41-kDa protein was present in fetal liver mitochondria but not in liver mitochondria from 5-day neonates. Thus disappearance of the 41-kDa protein coincided with the developmental decline in liver branched chain aminotransferase activity. Two-dimensional immunoblots of isolated BCATm immunocomplexes showed that the liver immunoreactive proteins were clearly different from the heart and kidney proteins which exhibited identical immunoblots. Investigation of BCATm in subcellular fractions prepared from different skeletal muscle fiber types revealed that branched chain aminotransferase is exclusively a mitochondrial enzyme in skeletal muscles. Although total detergent-extractable branched chain aminotransferase activity was largely independent of fiber type, branched chain aminotransferase activity and BCATm protein concentration were highest in mitochondria prepared from white gastrocnemius followed by mixed skeletal muscles with lowest activity and protein concentration found in soleus mitochondria. These quantitative differences in mitochondrial branched chain aminotransferase activity and enzyme protein content suggest there may be differential expression of BCATm in different muscle fiber types.
authors.affiliation: Department of Biochemistry, Wake Forest University, Bowman Gray School of Medicine, Winston-Salem, North Carolina 27157.
authors.forename: S M
authors.initials: SM
authors.lastname: Hutson
authors.forename: R
authors.initials: R
authors.lastname: Wallin
authors.forename: T R
authors.initials: TR
authors.lastname: Hall
mesh_terms: D000375:Aging; D000818:Animals; D000831:Animals, Newborn; D001921:Brain / Q000201:enzymology; D015180:Electrophoresis, Gel, Two-Dimensional; D004591:Electrophoresis, Polyacrylamide Gel; D015151:Immunoblotting; D007527:Isoenzymes / Q000032:analysis* / Q000302:isolation & purification; D007668:Kidney / Q000201:enzymology; D008099:Liver / Q000254:growth & development; D008928:Mitochondria / Q000201:enzymology*; D008929:Mitochondria, Heart / Q000201:enzymology; D008930:Mitochondria, Liver / Q000201:enzymology; D008970:Molecular Weight; D009132:Muscles / Q000201:enzymology*; D009928:Organ Specificity; D051381:Rats; D011919:Rats, Inbred Strains; D013270:Stomach / Q000201:enzymology; D013347:Subcellular Fractions / Q000201:enzymology; D000637:Transaminases / Q000032:analysis* / Q000378:metabolism
chemical_list: D007527:Isoenzymes; D000637:Transaminases; C019798:branched-chain-amino-acid transaminase