Visualization of purified fibronectin-transglutaminase complexes

Ellen LeMosy, H. P. Erickson, W. F. Beyer, J. T. Radek, J. M. Jeong, S. N.P. Murthy, L. Lorand

Research output: Contribution to journalArticle

45 Citations (Scopus)

Abstract

It has been reported previously (Turner, P. M., and Lorand, L. (1989) Biochemistry 28, 628-635) that human erythrocyte transglutaminase forms a noncovalent complex with human plasma fibronectin near its collagen-binding domain. In the present study, we show by nondenaturing electrophoresis that guinea pig liver transglutaminase, similarly to the erythrocyte enzyme, forms a complex with human fibronectin. Studies of anisotropic shifts of fluorescein-labeled liver and erythrocyte transglutaminases, upon addition of fibronectin, indicated that both transglutaminases bind to fibronectin with a stoichiometry of about 2:1. Polymerization of fibrinogen by human erythrocyte transglutaminase was inhibited after complex formation with fibronectin. Complexes of fibronectin with either erythrocyte or liver transglutaminase were isolated by glycerol gradient zone sedimentation and examined by rotary shadowing electron microscopy. The globular transglutaminase could be readily identified binding to the thin fibronectin strand. The binding site for transglutaminase was within 5-10 nm of the N terminus of fibronectin, consistent with its proximity to the collagen-binding domain. Under some experimental conditions, the complex of fibronectin with erythrocyte transglutaminase appeared as a ring-shaped structure in which two transglutaminase molecules had probably dimerized. The molecular weight of the erythrocyte transglutaminase was determined by sedimentation equilibrium to be 71,440 ± 830.

Original languageEnglish (US)
Pages (from-to)7880-7885
Number of pages6
JournalJournal of Biological Chemistry
Volume267
Issue number11
StatePublished - Jan 1 1992
Externally publishedYes

Fingerprint

Transglutaminases
Fibronectins
Visualization
Erythrocytes
Liver
Sedimentation
Collagen
Plasma (human)
Biochemistry
Electrophoresis
Fluorescein
Stoichiometry
Polymerization
Glycerol
Fibrinogen
Electron microscopy
Electron Microscopy
Guinea Pigs
Molecular Weight
Molecular weight

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

LeMosy, E., Erickson, H. P., Beyer, W. F., Radek, J. T., Jeong, J. M., Murthy, S. N. P., & Lorand, L. (1992). Visualization of purified fibronectin-transglutaminase complexes. Journal of Biological Chemistry, 267(11), 7880-7885.

Visualization of purified fibronectin-transglutaminase complexes. / LeMosy, Ellen; Erickson, H. P.; Beyer, W. F.; Radek, J. T.; Jeong, J. M.; Murthy, S. N.P.; Lorand, L.

In: Journal of Biological Chemistry, Vol. 267, No. 11, 01.01.1992, p. 7880-7885.

Research output: Contribution to journalArticle

LeMosy, E, Erickson, HP, Beyer, WF, Radek, JT, Jeong, JM, Murthy, SNP & Lorand, L 1992, 'Visualization of purified fibronectin-transglutaminase complexes', Journal of Biological Chemistry, vol. 267, no. 11, pp. 7880-7885.
LeMosy E, Erickson HP, Beyer WF, Radek JT, Jeong JM, Murthy SNP et al. Visualization of purified fibronectin-transglutaminase complexes. Journal of Biological Chemistry. 1992 Jan 1;267(11):7880-7885.
LeMosy, Ellen ; Erickson, H. P. ; Beyer, W. F. ; Radek, J. T. ; Jeong, J. M. ; Murthy, S. N.P. ; Lorand, L. / Visualization of purified fibronectin-transglutaminase complexes. In: Journal of Biological Chemistry. 1992 ; Vol. 267, No. 11. pp. 7880-7885.
@article{d451e7b93ec74f54aa810c92e3638be0,
title = "Visualization of purified fibronectin-transglutaminase complexes",
abstract = "It has been reported previously (Turner, P. M., and Lorand, L. (1989) Biochemistry 28, 628-635) that human erythrocyte transglutaminase forms a noncovalent complex with human plasma fibronectin near its collagen-binding domain. In the present study, we show by nondenaturing electrophoresis that guinea pig liver transglutaminase, similarly to the erythrocyte enzyme, forms a complex with human fibronectin. Studies of anisotropic shifts of fluorescein-labeled liver and erythrocyte transglutaminases, upon addition of fibronectin, indicated that both transglutaminases bind to fibronectin with a stoichiometry of about 2:1. Polymerization of fibrinogen by human erythrocyte transglutaminase was inhibited after complex formation with fibronectin. Complexes of fibronectin with either erythrocyte or liver transglutaminase were isolated by glycerol gradient zone sedimentation and examined by rotary shadowing electron microscopy. The globular transglutaminase could be readily identified binding to the thin fibronectin strand. The binding site for transglutaminase was within 5-10 nm of the N terminus of fibronectin, consistent with its proximity to the collagen-binding domain. Under some experimental conditions, the complex of fibronectin with erythrocyte transglutaminase appeared as a ring-shaped structure in which two transglutaminase molecules had probably dimerized. The molecular weight of the erythrocyte transglutaminase was determined by sedimentation equilibrium to be 71,440 ± 830.",
author = "Ellen LeMosy and Erickson, {H. P.} and Beyer, {W. F.} and Radek, {J. T.} and Jeong, {J. M.} and Murthy, {S. N.P.} and L. Lorand",
year = "1992",
month = "1",
day = "1",
language = "English (US)",
volume = "267",
pages = "7880--7885",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology Inc.",
number = "11",

}

TY - JOUR

T1 - Visualization of purified fibronectin-transglutaminase complexes

AU - LeMosy, Ellen

AU - Erickson, H. P.

AU - Beyer, W. F.

AU - Radek, J. T.

AU - Jeong, J. M.

AU - Murthy, S. N.P.

AU - Lorand, L.

PY - 1992/1/1

Y1 - 1992/1/1

N2 - It has been reported previously (Turner, P. M., and Lorand, L. (1989) Biochemistry 28, 628-635) that human erythrocyte transglutaminase forms a noncovalent complex with human plasma fibronectin near its collagen-binding domain. In the present study, we show by nondenaturing electrophoresis that guinea pig liver transglutaminase, similarly to the erythrocyte enzyme, forms a complex with human fibronectin. Studies of anisotropic shifts of fluorescein-labeled liver and erythrocyte transglutaminases, upon addition of fibronectin, indicated that both transglutaminases bind to fibronectin with a stoichiometry of about 2:1. Polymerization of fibrinogen by human erythrocyte transglutaminase was inhibited after complex formation with fibronectin. Complexes of fibronectin with either erythrocyte or liver transglutaminase were isolated by glycerol gradient zone sedimentation and examined by rotary shadowing electron microscopy. The globular transglutaminase could be readily identified binding to the thin fibronectin strand. The binding site for transglutaminase was within 5-10 nm of the N terminus of fibronectin, consistent with its proximity to the collagen-binding domain. Under some experimental conditions, the complex of fibronectin with erythrocyte transglutaminase appeared as a ring-shaped structure in which two transglutaminase molecules had probably dimerized. The molecular weight of the erythrocyte transglutaminase was determined by sedimentation equilibrium to be 71,440 ± 830.

AB - It has been reported previously (Turner, P. M., and Lorand, L. (1989) Biochemistry 28, 628-635) that human erythrocyte transglutaminase forms a noncovalent complex with human plasma fibronectin near its collagen-binding domain. In the present study, we show by nondenaturing electrophoresis that guinea pig liver transglutaminase, similarly to the erythrocyte enzyme, forms a complex with human fibronectin. Studies of anisotropic shifts of fluorescein-labeled liver and erythrocyte transglutaminases, upon addition of fibronectin, indicated that both transglutaminases bind to fibronectin with a stoichiometry of about 2:1. Polymerization of fibrinogen by human erythrocyte transglutaminase was inhibited after complex formation with fibronectin. Complexes of fibronectin with either erythrocyte or liver transglutaminase were isolated by glycerol gradient zone sedimentation and examined by rotary shadowing electron microscopy. The globular transglutaminase could be readily identified binding to the thin fibronectin strand. The binding site for transglutaminase was within 5-10 nm of the N terminus of fibronectin, consistent with its proximity to the collagen-binding domain. Under some experimental conditions, the complex of fibronectin with erythrocyte transglutaminase appeared as a ring-shaped structure in which two transglutaminase molecules had probably dimerized. The molecular weight of the erythrocyte transglutaminase was determined by sedimentation equilibrium to be 71,440 ± 830.

UR - http://www.scopus.com/inward/record.url?scp=0026655913&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0026655913&partnerID=8YFLogxK

M3 - Article

VL - 267

SP - 7880

EP - 7885

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 11

ER -