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Characterization of cDNA for murine tripeptidyl-peptidase II reveals alternative splicing

B Tomkinson1

  • 1Department of Veterinary Medical Chemistry, Swedish University of Agricultural Sciences, Uppsala.

The Biochemical Journal
|December 1, 1994
PubMed

Insights

Cloning murine tripeptidyl-peptidase II (TPP II) revealed high sequence identity with the human enzyme. This suggests the entire TPP II subunit is functionally important, with a unique domain structure compared to other subtilases.

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Enzymology

Background:

  • Tripeptidyl-peptidase II (TPP II) is a cytosolic exopeptidase.
  • TPP II possesses a catalytic active site characteristic of subtilisin-type proteases.

Purpose of the Study:

  • To clone the cDNA encoding murine TPP II.
  • To analyze the deduced amino acid sequence and compare it with human TPP II.
  • To investigate the structural and functional implications of TPP II's sequence.

Main Methods:

  • cDNA library screening from murine mastocytoma.
  • 5'-RACE (rapid amplification of cDNA ends) for sequence isolation.
  • Sequence alignment with subtilisin-like serine peptidases.

Main Results:

  • Isolated 4611 bp of murine TPP II cDNA, including the complete coding region.
  • Deduced amino acid sequence showed 96% identity to human TPP II.
  • Identified a catalytic triad (Asp44, His264, Ser449) and a unique ~200 amino acid domain.
  • Observed alternative polyadenylation signals and evidence of alternative splicing.

Conclusions:

  • Murine and human TPP II share high sequence identity, indicating conserved functional importance of the entire subunit.
  • TPP II exhibits a distinct domain structure compared to other subtilases.
  • The study highlights mRNA processing variations, including alternative splicing and polyadenylation, in TPP II expression.

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