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Identification and Kinetics Analysis of a Novel Heparin-binding Site (KEDK) in Human Tenascin-C

Cited 13 time in Web of Science Cited 14 time in Scopus
Authors
Jang, Jun-Hyeong; Hwang, Jung-Hee; Chung, Chong-Pyoung; Choung, Pill-Hoon
Issue Date
2004-06
Publisher
The American Society for Biochemistry and Molecular Biology
Citation
Journal of Biological Chemistry, Vol. 279, No. 24, pp. 25562–25566, 2004
Abstract
The interaction between tenascin-C (TN-C), a multi-subunit extracellular matrix protein, and heparin was examined using a surface plasmon resonance-based technique on a Biacore system. The aims of the present study were to examine the affinity of fibronectin type III repeats of TN-C fragments (TNIII) for heparin, to investigate the role of the TNIII4 domains in the binding of TN-C to heparin, and to delineate a sequence of amino acids within the TNIII4 domain, which mediates cooperative heparin binding. At a physiological salt concentration, and pH 7.4, TNIII3–5 binds to heparin with high affinity (KD = 30 nm). However, a major heparin-binding site in TNIII5 produces a modest affinity binding at a KD near 4 μm, and a second site in TNIII4 enhances the binding by several orders of magnitude, although it was far too weak to produce an observable binding of TNIII4 by itself. Moreover, mutagenesis of the KEDK sequence in the TNIII4 domain resulted in the significant reduction of heparin-binding affinity. In addition, residues in the KEDK sequences are conserved in TN-C throughout mammalian evolution. Thus the structure-based sequence alignment, mutagenesis, and sequence conservation data together reveal a KEDK sequence in TNIII4 suggestive of a minor heparin-binding site. Finally, we demonstrate that TNIII4 contains binding sites for heparin sulfate proteoglycan and enhances the heparin sulfate proteoglycan-dependent human gingival fibroblast adhesion to TNIII5, thus providing the biological significance of heparin-binding site of TNIII4. These results suggest that the heparin-binding sites may traverse TNIII4–5 and thus require KEDK in TNIII4 for optimal heparin-binding.
ISSN
0021-9258
Language
English
URI
http://hdl.handle.net/10371/20264
DOI
https://doi.org/10.1074/jbc.M403170200
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College of Dentistry/School of Dentistry (치과대학/치의학대학원)Dept. of Dentistry (치의학과)Journal Papers (저널논문_치의학과)
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