Interaction with Heparin and Matrix Metalloproteinase 2 Cleavage Expose a Cryptic Anti-adhesive Site of Fibronectin
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- 作者：Kazuo Watanabe ; Hiroshi Takahashi ; Yoshiko Habu ; Naoko Kamiya-Kubushiro ; Sadahiro Kamiya ; Hiroshi Nakamura ; Hirofumi Yajima ; Tadahiro Ishii ; Takashi Katayama ; Kaoru Miyazaki ; and Fumio Fukai
- 刊名：Biochemistry
- 出版年：2000
- 出版时间：June 20, 2000
- 年：2000
- 卷：39
- 期：24
- 页码：7138 - 7144
- 全文大小：180K
- 年卷期：v.39,no.24(June 20, 2000)
- ISSN：1520-4995
文摘
We recently found that fibronectin (FN) had a functional site [YTIYVIAL sequence in theheparin-binding domain 2 (Hep 2)] that was capable of suppressing the integrin-mediated cell adhesionto extracellular matrix. However, our results also indicated that this anti-adhesive site seemed to be usuallyburied within the Hep 2 domain structure because of its hydrophobic nature, raising a question as to thephysiological significance of the cryptic anti-adhesive activity of FN. The present study demonstratesthat the cryptic anti-adhesive activity can be exposed through the physiological processes. A 30-kDachymotryptic FN fragment derived from Hep 2 domain (Hep 2 fragment), which had no effect on adhesionof MSV-transformed nonproducer 3T3 cell line (KN78) to FN, expressed the anti-adhesive activity aftertreatment with 6 M urea. Light scattering and circular dichroism measurements showed that the ureatreatment induced the conformational change of the Hep 2 fragment from a more compact form to anunfolded one. Incubation of the Hep 2 fragment with heparin also induced similar conformational changesand expression of anti-adhesive activity. Additionally, both the urea and heparin treatments made theHep 2 fragment and intact FN much more accessible to the polyclonal antibody (
III14A), with arecognition site near the anti-adhesive site of FN. Specific cleavage of either the Hep 2 fragment or intactFN by matrix metalloproteinase 2 (MMP-2) released a 10-kDa fragment with the anti-adhesive activity,which was shown to have the exposed anti-adhesive site on the amino-terminal region. Thus, the crypticanti-adhesive activity of FN can be expressed upon conformational change and proteolytic cleavage ofHep 2 domain.
III14A), with arecognition site near the anti-adhesive site of FN. Specific cleavage of either the Hep 2 fragment or intactFN by matrix metalloproteinase 2 (MMP-2) released a 10-kDa fragment with the anti-adhesive activity,which was shown to have the exposed anti-adhesive site on the amino-terminal region. Thus, the crypticanti-adhesive activity of FN can be expressed upon conformational change and proteolytic cleavage ofHep 2 domain.