Demonstration of the extrinsic coagulation pathway in teleostei

Identification of zebrafish coagulation factor VII

John Sheehan, Michael Templer, Michael Gregory, Ravikumar Hanumanthaiah, Dean Troyer, Thao Phan, Bharath Thankavel, Pudur Jagadeeswaran

Research output: Contribution to journalArticle

57 Citations (Scopus)

Abstract

It is not known whether the mammalian mechanism of coagulation initiation is conserved in fish. Identification of factor VII is critical in providing evidence for such a mechanism. A cDNA was cloned from a zebrafish (teleost) library that predicted a protein with sequence similarity to human factor VII. Factor VII was shown to be present in zebrafish blood and liver by Western blot analysis and immunohistochemistry. Immunodepletion of factor VII from zebrafish plasma selectively inhibited thromboplastin-triggered thrombin generation. Heterologous expression of zebrafish factor VII demonstrated a secreted protein (50 kDa) that reconstituted thromboplastin-triggered thrombin generation in immunodepleted zebrafish plasma. These results suggest conservation of the extrinsic coagulation pathway between zebrafish and humans and add credence to the zebrafish as a model for mammalian hemostasis. The structure of zebrafish factor VIIa predicted by homology modeling was consistent with the overall three-dimensional structure of human factor VIIa. However, amino acid disparities were found in the epidermal growth factor-2/serine protease regions that are present in the human tissue factor-factor VIIa contact surface, suggesting a structural basis for the species specificity of this interaction. In addition, zebrafish factor VII demonstrates that the Gla-EGF-EGF-SP domain structure, which is common to coagulation factors VII, IX, X, and protein C, was present before the radiation of the teleosts from the tetrapods. Identification of zebrafish factor VII significantly narrows the evolutionary window for development of the vertebrate coagulation cascade and provides insight into the structural basis for species specificity in the tissue factor-factor VIIa interaction.

Original languageEnglish (US)
Pages (from-to)8768-8773
Number of pages6
JournalProceedings of the National Academy of Sciences of the United States of America
Volume98
Issue number15
DOIs
StatePublished - Jul 17 2001

Fingerprint

Factor VII
Zebrafish
Factor VIIa
Thromboplastin
Epidermal Growth Factor
Species Specificity
Thrombin
Factor IX
Serine Proteases
Protein C
Hemostasis
Libraries
Vertebrates
Fishes
Proteins
Complementary DNA
Western Blotting
Immunohistochemistry
Radiation
Amino Acids

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Demonstration of the extrinsic coagulation pathway in teleostei : Identification of zebrafish coagulation factor VII. / Sheehan, John; Templer, Michael; Gregory, Michael; Hanumanthaiah, Ravikumar; Troyer, Dean; Phan, Thao; Thankavel, Bharath; Jagadeeswaran, Pudur.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 98, No. 15, 17.07.2001, p. 8768-8773.

Research output: Contribution to journalArticle

Sheehan, John ; Templer, Michael ; Gregory, Michael ; Hanumanthaiah, Ravikumar ; Troyer, Dean ; Phan, Thao ; Thankavel, Bharath ; Jagadeeswaran, Pudur. / Demonstration of the extrinsic coagulation pathway in teleostei : Identification of zebrafish coagulation factor VII. In: Proceedings of the National Academy of Sciences of the United States of America. 2001 ; Vol. 98, No. 15. pp. 8768-8773.
@article{b175ba95390846f78a8b47eb27a94951,
title = "Demonstration of the extrinsic coagulation pathway in teleostei: Identification of zebrafish coagulation factor VII",
abstract = "It is not known whether the mammalian mechanism of coagulation initiation is conserved in fish. Identification of factor VII is critical in providing evidence for such a mechanism. A cDNA was cloned from a zebrafish (teleost) library that predicted a protein with sequence similarity to human factor VII. Factor VII was shown to be present in zebrafish blood and liver by Western blot analysis and immunohistochemistry. Immunodepletion of factor VII from zebrafish plasma selectively inhibited thromboplastin-triggered thrombin generation. Heterologous expression of zebrafish factor VII demonstrated a secreted protein (50 kDa) that reconstituted thromboplastin-triggered thrombin generation in immunodepleted zebrafish plasma. These results suggest conservation of the extrinsic coagulation pathway between zebrafish and humans and add credence to the zebrafish as a model for mammalian hemostasis. The structure of zebrafish factor VIIa predicted by homology modeling was consistent with the overall three-dimensional structure of human factor VIIa. However, amino acid disparities were found in the epidermal growth factor-2/serine protease regions that are present in the human tissue factor-factor VIIa contact surface, suggesting a structural basis for the species specificity of this interaction. In addition, zebrafish factor VII demonstrates that the Gla-EGF-EGF-SP domain structure, which is common to coagulation factors VII, IX, X, and protein C, was present before the radiation of the teleosts from the tetrapods. Identification of zebrafish factor VII significantly narrows the evolutionary window for development of the vertebrate coagulation cascade and provides insight into the structural basis for species specificity in the tissue factor-factor VIIa interaction.",
author = "John Sheehan and Michael Templer and Michael Gregory and Ravikumar Hanumanthaiah and Dean Troyer and Thao Phan and Bharath Thankavel and Pudur Jagadeeswaran",
year = "2001",
month = "7",
day = "17",
doi = "10.1073/pnas.131109398",
language = "English (US)",
volume = "98",
pages = "8768--8773",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
number = "15",

}

TY - JOUR

T1 - Demonstration of the extrinsic coagulation pathway in teleostei

T2 - Identification of zebrafish coagulation factor VII

AU - Sheehan, John

AU - Templer, Michael

AU - Gregory, Michael

AU - Hanumanthaiah, Ravikumar

AU - Troyer, Dean

AU - Phan, Thao

AU - Thankavel, Bharath

AU - Jagadeeswaran, Pudur

PY - 2001/7/17

Y1 - 2001/7/17

N2 - It is not known whether the mammalian mechanism of coagulation initiation is conserved in fish. Identification of factor VII is critical in providing evidence for such a mechanism. A cDNA was cloned from a zebrafish (teleost) library that predicted a protein with sequence similarity to human factor VII. Factor VII was shown to be present in zebrafish blood and liver by Western blot analysis and immunohistochemistry. Immunodepletion of factor VII from zebrafish plasma selectively inhibited thromboplastin-triggered thrombin generation. Heterologous expression of zebrafish factor VII demonstrated a secreted protein (50 kDa) that reconstituted thromboplastin-triggered thrombin generation in immunodepleted zebrafish plasma. These results suggest conservation of the extrinsic coagulation pathway between zebrafish and humans and add credence to the zebrafish as a model for mammalian hemostasis. The structure of zebrafish factor VIIa predicted by homology modeling was consistent with the overall three-dimensional structure of human factor VIIa. However, amino acid disparities were found in the epidermal growth factor-2/serine protease regions that are present in the human tissue factor-factor VIIa contact surface, suggesting a structural basis for the species specificity of this interaction. In addition, zebrafish factor VII demonstrates that the Gla-EGF-EGF-SP domain structure, which is common to coagulation factors VII, IX, X, and protein C, was present before the radiation of the teleosts from the tetrapods. Identification of zebrafish factor VII significantly narrows the evolutionary window for development of the vertebrate coagulation cascade and provides insight into the structural basis for species specificity in the tissue factor-factor VIIa interaction.

AB - It is not known whether the mammalian mechanism of coagulation initiation is conserved in fish. Identification of factor VII is critical in providing evidence for such a mechanism. A cDNA was cloned from a zebrafish (teleost) library that predicted a protein with sequence similarity to human factor VII. Factor VII was shown to be present in zebrafish blood and liver by Western blot analysis and immunohistochemistry. Immunodepletion of factor VII from zebrafish plasma selectively inhibited thromboplastin-triggered thrombin generation. Heterologous expression of zebrafish factor VII demonstrated a secreted protein (50 kDa) that reconstituted thromboplastin-triggered thrombin generation in immunodepleted zebrafish plasma. These results suggest conservation of the extrinsic coagulation pathway between zebrafish and humans and add credence to the zebrafish as a model for mammalian hemostasis. The structure of zebrafish factor VIIa predicted by homology modeling was consistent with the overall three-dimensional structure of human factor VIIa. However, amino acid disparities were found in the epidermal growth factor-2/serine protease regions that are present in the human tissue factor-factor VIIa contact surface, suggesting a structural basis for the species specificity of this interaction. In addition, zebrafish factor VII demonstrates that the Gla-EGF-EGF-SP domain structure, which is common to coagulation factors VII, IX, X, and protein C, was present before the radiation of the teleosts from the tetrapods. Identification of zebrafish factor VII significantly narrows the evolutionary window for development of the vertebrate coagulation cascade and provides insight into the structural basis for species specificity in the tissue factor-factor VIIa interaction.

UR - http://www.scopus.com/inward/record.url?scp=0035902566&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0035902566&partnerID=8YFLogxK

U2 - 10.1073/pnas.131109398

DO - 10.1073/pnas.131109398

M3 - Article

VL - 98

SP - 8768

EP - 8773

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 15

ER -