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 language | English |
|---|---|
| Pages (from-to) | 477-481 |
| Number of pages | 5 |
| Journal | Biological and Pharmaceutical Bulletin |
| Volume | 23 |
| Issue number | 4 |
| Publication status | Published - 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
Influence of N-terminal peptide and oligosaccharide on the clearance of t-PA. / Aoki, Shoichi; Shimizu, Norihide; Koyama, Jun Ichi; Kato, Yoshiko; Kitagawa, Masaru; Okumura, Kazuo; Tanigawara, Yusuke.
In: Biological and Pharmaceutical Bulletin, Vol. 23, No. 4, 04.2000, p. 477-481.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Influence of N-terminal peptide and oligosaccharide on the clearance of t-PA
AU - Aoki, Shoichi
AU - Shimizu, Norihide
AU - Koyama, Jun Ichi
AU - Kato, Yoshiko
AU - Kitagawa, Masaru
AU - Okumura, Kazuo
AU - Tanigawara, Yusuke
PY - 2000/4
Y1 - 2000/4
N2 - 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.
AB - 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.
KW - Gln117 t-PA
KW - L-chain
KW - S-chain
KW - Tissue plasminogen activator
KW - Variant
UR - http://www.scopus.com/inward/record.url?scp=0034014690&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=0034014690&partnerID=8YFLogxK
M3 - Article
C2 - 10784431
AN - SCOPUS:0034014690
VL - 23
SP - 477
EP - 481
JO - Biological and Pharmaceutical Bulletin
JF - Biological and Pharmaceutical Bulletin
SN - 0918-6158
IS - 4
ER -