Influence of N-terminal peptide and oligosaccharide on the clearance of t-PA

Shoichi Aoki, Norihide Shimizu, Jun Ichi Koyama, Yoshiko Kato, Masaru Kitagawa, Kazuo Okumura, Yusuke Tanigawara

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

We have studied the influence of Gly-Ala-Arg peptide at the N-terminus and the oligosaccharide at Asn184 on the clearance of tissue plasminogen activator (t-PA). In order to intensify the influence of these structural features, Gln117 t-PA, which is a mutant tissue plasminogen activator (mt-PA) expressed in mouse C127 cells, was used for the investigation. It is altered to remove a high mannose type oligosaccharide by the mutation of an amino acid from Asn117 to Gln. We isolated 4 variants of Gln117 t-PA by cation- exchange chromatography, which are abbreviated as S-I, S-II, L-I and L-II. These variants originated from the heterogeneity of the peptide chains (S- chain, 527 amino acids, L-chain, 530 amino acids) and oligosaccharide (Type I, 2 oligosaccharides, Type II, 1 oligosaccharide). Pharmacokinetics of these variants were investigated after single intravenous administration to male rats at a dose of 250 μg/kg. Significant differences in pharmacokinetic parameters were observed among these variants, but there was no considerable difference in fibrin clot lysis time (FCLT) activity. Gly-Ala-Arg peptide at the N-terminus increased the CL(t), whereas the oligosaccharide at Asn184 decreased the CL(t). Moreover, the effects of the N-terminal peptide and the oligosaccharide on the CL(t) were independent of each other. Our study with Gln117 t-PA revealed the role of the N-terminal peptide found in the L-chain produced during the processing of t-PA precursor.

Original languageEnglish
Pages (from-to)477-481
Number of pages5
JournalBiological and Pharmaceutical Bulletin
Volume23
Issue number4
Publication statusPublished - 2000 Apr

    Fingerprint

Keywords

  • Gln117 t-PA
  • L-chain
  • S-chain
  • Tissue plasminogen activator
  • Variant

ASJC Scopus subject areas

  • Molecular Medicine
  • Pharmacology, Toxicology and Pharmaceutics(all)

Cite this

Aoki, S., Shimizu, N., Koyama, J. I., Kato, Y., Kitagawa, M., Okumura, K., & Tanigawara, Y. (2000). Influence of N-terminal peptide and oligosaccharide on the clearance of t-PA. Biological and Pharmaceutical Bulletin, 23(4), 477-481.