May 2004
Volume 45, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2004
Characterization of transforming growth factor beta induced protein (TGFBIp) purified from normal human and porcine corneas
J.J. Enghild; R.B. Andersen; Z. Valnickova; I.B. Thogersen; C.J. Hedegaard; T. Kristensen; G.K. Klintworth; T. Moller–Pedersen
Author Affiliations & Notes
  • J.J. Enghild
    Department of Molecular Biology, Aarhus University, Aarhus, Denmark
  • R.B. Andersen
    Department of Molecular Biology, Aarhus University, Aarhus, Denmark
  • Z. Valnickova
    Department of Molecular Biology, Aarhus University, Aarhus, Denmark
  • I.B. Thogersen
    Department of Molecular Biology, Aarhus University, Aarhus, Denmark
  • C.J. Hedegaard
    Department of Molecular Biology, Aarhus University, Aarhus, Denmark
  • T. Kristensen
    Department of Molecular Biology, Aarhus University, Aarhus, Denmark
  • G.K. Klintworth
    Department of Pathology, Duke University Medical Center, Durham, NC
  • T. Moller–Pedersen
    Department of Ophthalmology, Aarhus University Hospital, Aarhus, Denmark
  • Footnotes
    Commercial Relationships  J.J. Enghild, None; R.B. Andersen, None; Z. Valnickova, None; I.B. Thogersen, None; C.J. Hedegaard, None; T. Kristensen, None; G.K. Klintworth, None; T. Moller–Pedersen, None.
  • Footnotes
    Support  NIH Grant RO1–EY12712
Investigative Ophthalmology & Visual Science May 2004, Vol.45, 3840. doi:
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      J.J. Enghild, R.B. Andersen, Z. Valnickova, I.B. Thogersen, C.J. Hedegaard, T. Kristensen, G.K. Klintworth, T. Moller–Pedersen; Characterization of transforming growth factor beta induced protein (TGFBIp) purified from normal human and porcine corneas . Invest. Ophthalmol. Vis. Sci. 2004;45(13):3840.

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Abstract

Abstract: : Purpose: To characterize TGFBIp (also termed BIGH3) isolated from normal human and porcine corneas and to examine corneal immunoreactivity to antibodies raised against central domains versus C–terminal fragments of TGFBIp. Methods: TGFBIp was purified from 20 human and 200 porcine corneas using standard chromatographic techniques under non–denaturing conditions and characterized by tandem mass spectrometry. Post–translational modifications were characterized by enzymatic fragmentation of purified TGFBIp. Specific rabbit antisera were raised against both central domains (amino acid residues 134 to 500) and C–terminal fragments (amino acids 648 to 683) of human TGFBIp. Five normal human corneas were processed for immunohistology. Results: Both human and porcine TGFBIp are 68 kDa monomers that are truncated at amino acid 647, three amino acid residues after the integrin binding RGD–sequence. Only small amounts of full–length TGFBIp exists in corneal extracts. Neither human nor porcine TGFBIp have posttranslational additions. A fraction of TGFBIp was covalently associated with insoluble extracellular matrix components. This interaction was disrupted following reduction, suggesting that binding was mediated by a disulfide bridge. The human cornea shows strong immunoreactivity towards central domains of TGFBIp, both inside the cells and in the extracellular matrix. By contrast, a much weaker and predominantly intracellular immunoreactivity was detected towards the C–terminal domains of TGFBIp, suggesting that the C–terminal fragment is cleaved as the protein leaves the cells. Conclusions: Normal human and porcine corneas contain abundant amounts of extracellular TGFBIp that exists predominantly as 68 kDa monomers following C–terminal clevage of amino acid residues 648 to 683.

Keywords: cornea: stroma and keratocytes • extracellular matrix • protein purification and characterization 
© 2004, The Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Permission to republish any abstract or part of an abstract in any form must be obtained in writing from the ARVO Office prior to publication.
Copyright © 2025 Association for Research in Vision and Ophthalmology.
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