A cDNA clone encoding enteropeptidase (EC 126.96.36.199), a key enzyme for the conversion of trypsinogen to trypsin, was isolated from a rat duodenal mucosa cDNA library. Sequence of the 3585 base pair clone predicted that enteropeptidase is synthesized as a single-chain precursor form, proenteropeptidase, consisting of 1058 amino acid residues with an internal signal sequence (51 residues) and is then processed into the mature enzyme consisting of three different peptide chains, i.e., mini, light and heavy chains, not the previously reported two-chain enzyme. The structure of enteropeptidase is relatively conserved among different species and the rat enteropeptidase is 24 and 39 amino acids longer than the porcine and human ones, respectively. Northern blot analysis of RNAs from normal rat tissues revealed that the enteropeptidase mRNA of around 4.4 kb in size was expressed only in the duodenal mucosa, and high proteolytic activity of the enzyme was detected in the proximal small intestine. Additional analysis of the RNAs by RT-PCR revealed that a low level of the mRNA was also expressed in the other parts of the small intestine, i.e., jejunum and ileum. These results indicate that the biosynthesis of enteropeptidase takes place mainly in the proximal small intestine, the duodenum, and the importance of the region in the physiology of intestinal protein digestion regulated by the enzyme is suggested. Furthermore a faint signal of the mRNA was also detected in the stomach, colon and brain in which the existence of trypsin-like serine proteases were reported. The significance of the low level expression of the gene is unclear, but the potential peptide-processing function of the enzyme in these tissues is also suggested.
|Number of pages||7|
|Journal||Biochemical and Biophysical Research Communications|
|Publication status||Published - 1996 Feb 27|
ASJC Scopus subject areas
- Molecular Biology
- Cell Biology