title: Alanine aminotransferase isoenzymes: molecular cloning and quantitative analysis of tissue expression in rats and serum elevation in liver toxicity.
abstract: The elevation of serum alanine aminotransferase (ALT) is regarded as an indicator of liver damage based on the presumption that ALT protein is specifically and abundantly expressed in the liver. However, ALT elevation is also observed in non-liver injury conditions (for example, muscle injury) and in apparently healthy people. Conversely, serum ALT activity is normal in many patients with confirmed liver diseases (for example, cirrhosis and hepatitis C infection). To improve the diagnostic value of the ALT assay and to understand the molecular basis for serum ALT changes in various pathophysiological conditions, we have cloned rat ALT isoenzyme ALT1 and ALT2 complementary DNAs (cDNAs), examined their tissue expressions at the messenger RNA and protein levels, and determined ALT1 and ALT 2 serum levels in response to liver damage in rodents. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis shows that ALT1 messenger RNA is widely distributed and mainly expressed in intestine, liver, fat tissues, colon, muscle, and heart, in the order of high to low expression level, whereas ALT2 gene expression is more restricted, mainly in liver, muscle, brain, and white adipose tissue. The tissue distribution pattern of ALT1 and ALT2 proteins largely agrees with their messenger RNA expression. Interestingly, hepatic ALT2 protein is approximately four times higher in male rats than in female rats. In addition, ALT isoenzymes distribute differentially at the subcellular level in that ALT1 is a cytoplasmic protein and ALT2 a mitochondrial protein, supporting bioinformatic prediction of mitochondrial localization of ALT2.
  
  
  CONCLUSIONS
  Using animal models of hepatoxicity induced by carbon tetrachloride and acetaminophen, we found that both serum ALT1 and ALT2 protein levels were significantly elevated and correlated with ALT activity, providing, for the first time, the molecular basis for the elevated total serum ALT activity.
authors.affiliation: Department of Medicine, Division of Endocrinology, University of Maryland School of Medicine, Baltimore, MD, USA.
authors.forename: Rong-Ze
authors.initials: RZ
authors.lastname: Yang
authors.forename: Soohyun
authors.initials: S
authors.lastname: Park
authors.forename: William J
authors.initials: WJ
authors.lastname: Reagan
authors.forename: Rick
authors.initials: R
authors.lastname: Goldstein
authors.forename: Shao
authors.initials: S
authors.lastname: Zhong
authors.forename: Michael
authors.initials: M
authors.lastname: Lawton
authors.forename: Francis
authors.initials: F
authors.lastname: Rajamohan
authors.forename: Kun
authors.initials: K
authors.lastname: Qian
authors.forename: Li
authors.initials: L
authors.lastname: Liu
authors.forename: Da-Wei
authors.initials: DW
authors.lastname: Gong
mesh_terms: D000410:Alanine Transaminase / Q000097:blood / Q000235:genetics*; D000595:Amino Acid Sequence; D000818:Animals; D003001:Cloning, Molecular; D017124:Conserved Sequence; D018076:DNA, Complementary / Q000235:genetics; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D007527:Isoenzymes / Q000097:blood / Q000235:genetics*; D008099:Liver / Q000201:enzymology / Q000473:pathology*; D008297:Male; D051379:Mice; D008930:Mitochondria, Liver / Q000201:enzymology; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D051381:Rats
chemical_list: D018076:DNA, Complementary; D007527:Isoenzymes; D000410:Alanine Transaminase