α-Amylase expressed in human liver is encoded by the AMY-2B gene identified in tumorous tissues

Iwao Koyama, Shin Ichi Komine, Nozomi Iino, Shigeru Hokari, Seiji Igarashi, David H. Alpers, Tsugikazu Komoda

Research output: Contribution to journalArticlepeer-review

12 Scopus citations

Abstract

Background: An α-Amylase in human liver is detected with an anti-human salivary amylase antibody, but the enzyme activity is very low. We previously found that the rat liver contained an amylase which differed from the enzyme of mice. In this study, we characterized the human liver amylases biochemically and immunohistochermically. Methods and Results: Although the amylase activity of human liver was much lower than that of rat, protein moiety and sugar chains of the human amylase were identified as similar to the rat liver enzyme with an anti-human salivary amylase antibody and by concanavalin A (Con A) affinity chromatography. Liver amylases from human and rat were the same size, 50 kDa, on Western blot analysis and had the same isoelectric points. The cytoplasm of hepatocytes was moderately stained immunohistochemically with the anti-human salivary amylase antibody. Intrahepatic bile ducts were also stained weak-to-moderately. RT-PCR, with a specific primer for the consensus sequence of human amylases, amplified a single 474-bp product from the human liver total RNA. The PCR product was sequenced and referred to the homology. Thirteen bases in the 434-bp fragment of the human liver amylase differed from the corresponding region of the AMY-1 gene transcript and the deduced amino acid sequence differed at five residues. The human liver amylase cDNA sequence was identical to the corresponding cDNA of the AMY-2B, which was known to expressed in tumorous tissues. In situ hybridization revealed the expression of AMY-2B mRNA in non-tumorous human liver. Conclusions: The present results suggest the possibility that a novel amylase detected in tumorous tissues and encoded by the AMY-2B gene is a liver-specific amylase expressed in the human liver.

Original languageEnglish
Pages (from-to)73-83
Number of pages11
JournalClinica Chimica Acta
Volume309
Issue number1
DOIs
StatePublished - 2001

Keywords

  • AMY-2B gene
  • Liver
  • Nucleotide sequence
  • RT-PCR
  • α-Amylase

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