A GPI-anchored co-receptor for tissue factor pathway inhibitor controls its intracellular trafficking and cell surface expression

S. A. Maroney, Anna Cunningham Edmondson, J. Ferrel, R. Hu, S. Haberichter, C. M. Mansbach, R. A. Brodsky, D. J. Dietzen, Alan E. Mast

Research output: Contribution to journalArticle

49 Citations (Scopus)

Abstract

Background: Tissue factor pathway inhibitor (TFPI) lacks a membrane attachment signal but it remains associated with the endothelial surface via its association with an, as yet, unidentified glycosyl phosphatidylinositol (GPI)-anchored coreceptor. Objectives/methods: Cellular trafficking of TFPI within aerolysin-resistant ECV304 and EA.hy926 cells, which do not express GPI-anchored proteins on their surface, was compared with their wild-type counterparts. Results and conclusions: Although aerolysin-resistant cells produce normal amounts of TFPI mRNA, TFPI is not expressed on the cell surface and total cellular TFPI is greatly decreased compared with wild-type cells. Additionally, normal, not increased, amounts of TFPI are secreted into conditioned media indicating that TFPI is degraded within the aerolysin-resistant cells. Confocal microscopy and studies using metabolic inhibitors demonstrate that aerolysin-resistant cells produce TFPI and transport it into the Golgi with subsequent degradation in lysosomes. The experimental results provide no evidence that cell surface FPI originates from secreted TFPI that binds back to aGPI-anchored protein. Instead, the data suggest that TFPI tightly, but reversibly, binds to a GPI anchored co-receptor in the ER/Golgi. The co-receptor then acts as a molecular chaperone for TFPI by trafficking it to the cell surface of wild-type cells or to lysosomes of aerolysin-resistant cells. TFPI that escapes co-receptor binding is secreted through the same pathway in both wild-type and aerolysin-resistant cells. The data provide a framework for understanding how TFPI is expressed on endothelium.

Original languageEnglish (US)
Pages (from-to)1114-1124
Number of pages11
JournalJournal of Thrombosis and Haemostasis
Volume4
Issue number5
DOIs
StatePublished - May 1 2006

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Glycosylphosphatidylinositols
Lysosomes
lipoprotein-associated coagulation inhibitor
Molecular Chaperones
Conditioned Culture Medium
Confocal Microscopy
Endothelium
aerolysin

Keywords

  • Aerolysin
  • Endothelial cells
  • GPI-anchor
  • Tissue factor pathway inhibitor

ASJC Scopus subject areas

  • Hematology

Cite this

A GPI-anchored co-receptor for tissue factor pathway inhibitor controls its intracellular trafficking and cell surface expression. / Maroney, S. A.; Edmondson, Anna Cunningham; Ferrel, J.; Hu, R.; Haberichter, S.; Mansbach, C. M.; Brodsky, R. A.; Dietzen, D. J.; Mast, Alan E.

In: Journal of Thrombosis and Haemostasis, Vol. 4, No. 5, 01.05.2006, p. 1114-1124.

Research output: Contribution to journalArticle

Maroney, S. A. ; Edmondson, Anna Cunningham ; Ferrel, J. ; Hu, R. ; Haberichter, S. ; Mansbach, C. M. ; Brodsky, R. A. ; Dietzen, D. J. ; Mast, Alan E. / A GPI-anchored co-receptor for tissue factor pathway inhibitor controls its intracellular trafficking and cell surface expression. In: Journal of Thrombosis and Haemostasis. 2006 ; Vol. 4, No. 5. pp. 1114-1124.
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T1 - A GPI-anchored co-receptor for tissue factor pathway inhibitor controls its intracellular trafficking and cell surface expression

AU - Maroney, S. A.

AU - Edmondson, Anna Cunningham

AU - Ferrel, J.

AU - Hu, R.

AU - Haberichter, S.

AU - Mansbach, C. M.

AU - Brodsky, R. A.

AU - Dietzen, D. J.

AU - Mast, Alan E.

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Y1 - 2006/5/1

N2 - Background: Tissue factor pathway inhibitor (TFPI) lacks a membrane attachment signal but it remains associated with the endothelial surface via its association with an, as yet, unidentified glycosyl phosphatidylinositol (GPI)-anchored coreceptor. Objectives/methods: Cellular trafficking of TFPI within aerolysin-resistant ECV304 and EA.hy926 cells, which do not express GPI-anchored proteins on their surface, was compared with their wild-type counterparts. Results and conclusions: Although aerolysin-resistant cells produce normal amounts of TFPI mRNA, TFPI is not expressed on the cell surface and total cellular TFPI is greatly decreased compared with wild-type cells. Additionally, normal, not increased, amounts of TFPI are secreted into conditioned media indicating that TFPI is degraded within the aerolysin-resistant cells. Confocal microscopy and studies using metabolic inhibitors demonstrate that aerolysin-resistant cells produce TFPI and transport it into the Golgi with subsequent degradation in lysosomes. The experimental results provide no evidence that cell surface FPI originates from secreted TFPI that binds back to aGPI-anchored protein. Instead, the data suggest that TFPI tightly, but reversibly, binds to a GPI anchored co-receptor in the ER/Golgi. The co-receptor then acts as a molecular chaperone for TFPI by trafficking it to the cell surface of wild-type cells or to lysosomes of aerolysin-resistant cells. TFPI that escapes co-receptor binding is secreted through the same pathway in both wild-type and aerolysin-resistant cells. The data provide a framework for understanding how TFPI is expressed on endothelium.

AB - Background: Tissue factor pathway inhibitor (TFPI) lacks a membrane attachment signal but it remains associated with the endothelial surface via its association with an, as yet, unidentified glycosyl phosphatidylinositol (GPI)-anchored coreceptor. Objectives/methods: Cellular trafficking of TFPI within aerolysin-resistant ECV304 and EA.hy926 cells, which do not express GPI-anchored proteins on their surface, was compared with their wild-type counterparts. Results and conclusions: Although aerolysin-resistant cells produce normal amounts of TFPI mRNA, TFPI is not expressed on the cell surface and total cellular TFPI is greatly decreased compared with wild-type cells. Additionally, normal, not increased, amounts of TFPI are secreted into conditioned media indicating that TFPI is degraded within the aerolysin-resistant cells. Confocal microscopy and studies using metabolic inhibitors demonstrate that aerolysin-resistant cells produce TFPI and transport it into the Golgi with subsequent degradation in lysosomes. The experimental results provide no evidence that cell surface FPI originates from secreted TFPI that binds back to aGPI-anchored protein. Instead, the data suggest that TFPI tightly, but reversibly, binds to a GPI anchored co-receptor in the ER/Golgi. The co-receptor then acts as a molecular chaperone for TFPI by trafficking it to the cell surface of wild-type cells or to lysosomes of aerolysin-resistant cells. TFPI that escapes co-receptor binding is secreted through the same pathway in both wild-type and aerolysin-resistant cells. The data provide a framework for understanding how TFPI is expressed on endothelium.

KW - Aerolysin

KW - Endothelial cells

KW - GPI-anchor

KW - Tissue factor pathway inhibitor

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