Design of variants of the second domain of urinary trypsin inhibitor (R-020) with increased factor Xa inhibitory activity.

Design of variants of the second domain of urinary trypsin inhibitor (R-020) with increased factor Xa inhibitory activity. Research Abstract Details 

Research Abstract Table of Contents

Jump to the:

Design of variants of the second domain of urinary trypsin inhibitor (R-020) with increased factor Xa inhibitory activity. Abstract Text:

A Nii,H Morishita,T Yamakawa,T Matsusue,J Hirose,T Miura,M Isaji,Y Horisawa,K Sugihara,T Kanamori,

The second domain (R-020) of human urinary trypsin inhibitor (UTI) exerts similar inhibitory activities on trypsin, alpha-chymotrypsin, leukocyte elastase, and plasmin to those of UTI itself, and additionally inhibits coagulation factor Xa (FXa) and plasma kallikrein, on both of which UTI has no inhibitory effect. In the present study, we attempted to increase this FXa-inhibitory activity by modeling the structure of R-020-FXa complex and substituting one or two amino acids in R-020 using recombinant DNA technology. Molecular modeling of R-020 and FXa was performed with reference to X-ray analysis of the complex of bovine pancreatic trypsin inhibitor (BPTI) and bovine trypsin to determine the site of amino acid modification. The expression plasmids into which R-020 genes with base substitution were inserted were prepared and introduced into Escherichia coli to express R-020 variants. The resulting variants were purified and their enzyme inhibitory activities were measured. The FXa-inhibitory activity was increased in four variants with single amino acid substitution. With another four variants having two amino acid substitutions involving combinations of the above single amino acid substitutions, the FXa-inhibitory activity was further increased. Because the electrostatic interaction within R-020-FXa complex seemed stronger in these R-020 variants, this increase in FXa-inhibitory effect was speculated to be a consequence of more potent binding between the enzyme and the inhibitor.

Design of variants of the second domain of urinary trypsin inhibitor (R-020) with increased factor Xa inhibitory activity. Publishing Authors By Initials

A Nii,H Morishita,T Yamakawa,T Matsusue,J Hirose,T Miura,M Isaji,Y Horisawa,K Sugihara,T Kanamori,

For similar chemical actions and uses: pharmacologic actions: molecular mechanisms of pharmacological action: enzyme inhibitors: protease inhibitors: serine proteinase inhibitors: trypsin inhibitors research abstracts see: chemical actions and uses: pharmacologic actions: molecular mechanisms of pharmacological action: enzyme inhibitors: protease inhibitors: serine proteinase inhibitors: trypsin inhibitors research

PUBMED ID PMID:

MEDLINE DATE:

Design of variants of the second domain of urinary trypsin inhibitor (R-020) with increased factor Xa inhibitory activity. Journal Published:

PUBLICATION TYPE: Journal Article

Journal: Journal of biochemistry

VOLUME: 115

Page Numbers: 1107-12

Journal Abbreviation: J. Biochem.

ISSN: 0021-924X

DAY: 19

MONTH: Jun

YEAR: 1994

Design of variants of the second domain of urinary trypsin inhibitor (R-020) with increased factor Xa inhibitory activity. Information

Number of References:

LANGUAGE: eng

NlmUniqueID: 376600

Design of variants of the second domain of urinary trypsin inhibitor (R-020) with increased factor Xa inhibitory activity. Keywords Mesh Terms:

KEYWORDS: Trypsin Inhibitors

MESH TERMS: metabolism

Chemical & Substance for Abstract: Design of variants of the second domain of urinary trypsin inhibitor (R-020) with increased factor Xa inhibitory activity. Information

Substance Name: Factor Xa

Registry Number: EC 3.4.21.6

Grant and Affiliation Information for Design of variants of the second domain of urinary trypsin inhibitor (R-020) with increased factor Xa inhibitory activity.

AFFILIATION: Biosciences Research Laboratory, Mochida Pharmaceutical Co., Ltd., Tokyo.

Country: JAPAN

AGENCY:

GRANT:

ACRONYM:

MEDLINETA: J Biochem

REFSOURCE:

DATABASENAME:

ACCESSION NUMBER:

Number Hits: 0

Design of variants of the second domain of urinary trypsin inhibitor R-020 with increased factor Xa inhibitory activity Related Publications

• An easily constructed, inexpensive device for dot blotting.
• Dementia Care Mapping-Japanese version(DCM-J)as a research evaluation method for measuring quality of life among elderly patients with dementia: Reliability and validity of Well-being and Ill-being value.
• Design of variants of the second domain of urinary trypsin inhibitor (R-020) with increased factor Xa inhibitory activity.
• Earthquake physics and real-time seismology.
• Isolation and characterization of the human chromosomal gene for prostacyclin-stimulating factor.
• Pressure-driven perfusion culture microchamber array for a parallel drug cytotoxicity assay.
• Purification of hyaluronidase from human placenta.
• Robust boosting algorithm against mislabeling in multiclass problems.
• Simultaneous multicentric occurrence of early hepatocellular carcinoma in a patient with persistent alpha-fetoprotein elevation.
• Surface modification of polydimethylsiloxane with photo-grafted poly(ethylene glycol) for micropatterned protein adsorption and cell adhesion.
• The complete amino acid sequence of a major trypsin inhibitor from seeds of foxtail millet (Setaria italica).
• [About chemotherapy of Yoshida sarcoma, with special reference to its relation to wandering velocity of leukocytes.]
• [Measurement and correction of diagnostic X-ray spectra using a high resolution Schottky CdTe detector]