Scripps VIVO scripps research logo

  • Index
  • Log in
  • Home
  • People
  • Organizations
  • Research
  • Events
Search form

Reversible regulation of tissue factor-induced coagulation by glycosyl phosphatidylinositol-anchored tissue factor pathway inhibitor

Academic Article
uri icon
  • Overview
  • Research
  • Identity
  • Additional Document Info
  • View All
scroll to property group menus

Overview

authors

  • Ott, I.
  • Miyagi, Y.
  • Miyazaki, K.
  • Heeb, Mary Jo
  • Mueller, B. M.
  • Rao, L. V. M.
  • Ruf, Wolfram

publication date

  • 2000

journal

  • Arteriosclerosis, Thrombosis, and Vascular Biology  Journal

abstract

  • Endothelial and tumor cells synthesize tissue factor pathway inhibitor (TFPI-1), which regulates tissue factor (TF) function by TF. VIIa. Xa. TFPI-1 quaternary complex formation (where VIIa and Xa are coagulation factors) and by translocation of these complexes into glycosphingolipid-rich microdomains of the cell membrane. Recombinant TFPI-1 added exogenously to cells is targeted to a degradation pathway. This study analyzes whether quaternary complex formation with endogenous TFPI-1 results also in internalization and degradation. We demonstrate that endogenous TFPI-1 and recombinant TFPI-1 differ in their distribution on the cell surface. Recombinant TFPI-1 is found in phospholipid- and glycosphingolipid-rich membrane domains, whereas endogenous TFPI-1 preferentially localizes to glycosphingolipid-rich microdomains. On quaternary complex formation, endogenous TFPI-1 remains protease sensitive and accessible for antibodies on intact cells, demonstrating that it is not appreciably internalized. Rather, regulation of TF by TFPI-1 is restored within 12 hours, consistent with dissociation of quaternary complexes on the cell surface. Endogenous TFPI-1 can be released from the cell surface by phospholipase treatment, indicating that TFPI-1 either is a glycosyl phosphatidylinositol (GPI)-anchored protein or binds to a GPI-linked receptor. We demonstrate that expression of a recombinant GPI-anchored form of TFPI-1 targets TF. VIIa complexes to glycosphingolipid-rich membrane fractions. Thus, GPI anchoring of TFPI-1 is sufficient for regulation of TF. VIIa complex function by a pathway of reversible inhibition rather than internalization and degradation.

subject areas

  • Animals
  • Antithrombin III
  • Binding Sites
  • Blood Coagulation
  • CHO Cells
  • Cricetinae
  • Detergents
  • Down-Regulation
  • Factor VIIa
  • Factor X
  • Factor Xa
  • Glycosylphosphatidylinositols
  • Humans
  • Membrane Proteins
  • Peptides
  • Plant Proteins
  • Protein Structure, Tertiary
  • Recombinant Proteins
  • Solubility
  • Thromboplastin
  • Trypsin Inhibitors
scroll to property group menus

Research

keywords

  • Kunitz-type inhibitors
  • cell surface proteoglycans
  • coagulation cascade
  • glycosphingolipid-rich microdomains
scroll to property group menus

Identity

International Standard Serial Number (ISSN)

  • 1079-5642

PubMed ID

  • 10712416
scroll to property group menus

Additional Document Info

start page

  • 874

end page

  • 882

volume

  • 20

issue

  • 3

©2019 The Scripps Research Institute | Terms of Use | Powered by VIVO

  • About
  • Contact Us
  • Support