Inhibition of factor VIIa-tissue factor coagulation activity by a hybrid protein

Thomas J. Girard; Louise A. Macphail; Karen M. Likert; William F. Novotny; Joseph P. Miletich; George J. Broze

doi:10.1126/science.1972598

Inhibition of factor VIIa-tissue factor coagulation activity by a hybrid protein

Thomas J. Girard, Louise A. Macphail, Karen M. Likert, William F. Novotny, Joseph P. Miletich, George J. Broze

Department of Pediatrics

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65 Scopus citations

Abstract

Lipoprotein-associated coagulation inhibitor (LACI) appears to inhibit tissue factor (TF)-induced blood coagulation by forming a quaternary inhibitory complex containing factor Xa, LACI, factor VIIa, and TF. A genetically engineered hybrid protein consisting of the light chain of factor Xa and the first Kunitz-type inhibitor domain of LACI is shown to directly inhibit the activity of the factor VIIa-TF catalytic complex. Unlike inhibition of factor VIIa-TF activity by native LACI, inhibition by the hybrid protein is not dependent on factor Xa. In an assay of TF-induced coagulation, 50% TF inhibition occurs with hybrid protein at 35 nanograms per milliliter, whereas LACI at 2.5 micrograms per milliliter is required for an equivalent effect. γ-Carboxylation of glutamic acid residues in the factor Xa light chain portion of the hybrid protein is required for inhibitory activity, indicating that the first Kunitz-type domain of LACI alone is not sufficient for inhibition of factor VIIa-TF.

Original language	English
Pages (from-to)	1421-1424
Number of pages	4
Journal	Science
Volume	248
Issue number	4961
DOIs	https://doi.org/10.1126/science.1972598
State	Published - 1990

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title = "Inhibition of factor VIIa-tissue factor coagulation activity by a hybrid protein",

abstract = "Lipoprotein-associated coagulation inhibitor (LACI) appears to inhibit tissue factor (TF)-induced blood coagulation by forming a quaternary inhibitory complex containing factor Xa, LACI, factor VIIa, and TF. A genetically engineered hybrid protein consisting of the light chain of factor Xa and the first Kunitz-type inhibitor domain of LACI is shown to directly inhibit the activity of the factor VIIa-TF catalytic complex. Unlike inhibition of factor VIIa-TF activity by native LACI, inhibition by the hybrid protein is not dependent on factor Xa. In an assay of TF-induced coagulation, 50% TF inhibition occurs with hybrid protein at 35 nanograms per milliliter, whereas LACI at 2.5 micrograms per milliliter is required for an equivalent effect. γ-Carboxylation of glutamic acid residues in the factor Xa light chain portion of the hybrid protein is required for inhibitory activity, indicating that the first Kunitz-type domain of LACI alone is not sufficient for inhibition of factor VIIa-TF.",

author = "Girard, {Thomas J.} and Macphail, {Louise A.} and Likert, {Karen M.} and Novotny, {William F.} and Miletich, {Joseph P.} and Broze, {George J.}",

year = "1990",

doi = "10.1126/science.1972598",

language = "English",

volume = "248",

pages = "1421--1424",

journal = "Science",

issn = "0036-8075",

number = "4961",

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T1 - Inhibition of factor VIIa-tissue factor coagulation activity by a hybrid protein

AU - Girard, Thomas J.

AU - Macphail, Louise A.

AU - Likert, Karen M.

AU - Novotny, William F.

AU - Miletich, Joseph P.

AU - Broze, George J.

PY - 1990

Y1 - 1990

N2 - Lipoprotein-associated coagulation inhibitor (LACI) appears to inhibit tissue factor (TF)-induced blood coagulation by forming a quaternary inhibitory complex containing factor Xa, LACI, factor VIIa, and TF. A genetically engineered hybrid protein consisting of the light chain of factor Xa and the first Kunitz-type inhibitor domain of LACI is shown to directly inhibit the activity of the factor VIIa-TF catalytic complex. Unlike inhibition of factor VIIa-TF activity by native LACI, inhibition by the hybrid protein is not dependent on factor Xa. In an assay of TF-induced coagulation, 50% TF inhibition occurs with hybrid protein at 35 nanograms per milliliter, whereas LACI at 2.5 micrograms per milliliter is required for an equivalent effect. γ-Carboxylation of glutamic acid residues in the factor Xa light chain portion of the hybrid protein is required for inhibitory activity, indicating that the first Kunitz-type domain of LACI alone is not sufficient for inhibition of factor VIIa-TF.

AB - Lipoprotein-associated coagulation inhibitor (LACI) appears to inhibit tissue factor (TF)-induced blood coagulation by forming a quaternary inhibitory complex containing factor Xa, LACI, factor VIIa, and TF. A genetically engineered hybrid protein consisting of the light chain of factor Xa and the first Kunitz-type inhibitor domain of LACI is shown to directly inhibit the activity of the factor VIIa-TF catalytic complex. Unlike inhibition of factor VIIa-TF activity by native LACI, inhibition by the hybrid protein is not dependent on factor Xa. In an assay of TF-induced coagulation, 50% TF inhibition occurs with hybrid protein at 35 nanograms per milliliter, whereas LACI at 2.5 micrograms per milliliter is required for an equivalent effect. γ-Carboxylation of glutamic acid residues in the factor Xa light chain portion of the hybrid protein is required for inhibitory activity, indicating that the first Kunitz-type domain of LACI alone is not sufficient for inhibition of factor VIIa-TF.

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U2 - 10.1126/science.1972598

DO - 10.1126/science.1972598

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AN - SCOPUS:0025291094

SN - 0036-8075

VL - 248

SP - 1421

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JO - Science

JF - Science

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ER -

Inhibition of factor VIIa-tissue factor coagulation activity by a hybrid protein

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