The reduced, denatured somatomedin B domain of vitronectin refolds into a stable, biologically active molecule

Biochemistry
Yuichi KamikuboD J Loskutoff

Abstract

The high-affinity binding site in human vitronectin (VN) for plasminogen activator inhibitor-1 (PAI-1) has been localized to the NH(2)-terminal cysteine-rich somatomedin B (SMB) domain (residues 1-44). A number of published structural and biochemical studies show conflicting results for the disulfide bonding pattern and the overall fold of the SMB domain, possibly because this domain may undergo disulfide shuffling and/or conformational changes during handling. Here we show that bacterially expressed recombinant SMB (rSMB) can be refolded to a single form that shows maximal activity in binding to PAI-1 and to a conformation-dependent monoclonal antibody (mAb 153). The oxidative refolding pathway of rSMB can be followed in the presence of glutathione redox buffers. This approach allowed the isolation and analysis of a number of intermediate folding species and of the final stably folded species at equilibrium. Competitive surface plasmon resonance analysis demonstrated that the stably refolded rSMB regained biological activity since it bound efficiently to PAI-1 and to mAb 153. In contrast, none of the folding intermediates bound to PAI-1 or to mAb 153. We also show by NMR analysis that the stably refolded rSMB is identical to t...Continue Reading

References

Nov 1, 1995·Journal of Biomolecular NMR·F DelaglioA Bax
Feb 1, 1996·Journal of Molecular Graphics·R KoradiK Wüthrich
Jun 4, 1998·Thrombosis Research·K T Preissner, D Seiffert
Jul 10, 1999·The International Journal of Biochemistry & Cell Biology·I SchvartzS Shaltiel
Aug 19, 2000·Cellular and Molecular Life Sciences : CMLS·P A AndreasenH H Petersen
May 23, 2002·The Journal of Biological Chemistry·Yu-ichi KamikuboDavid J Loskutoff
Sep 1, 1994·Journal of Biomolecular NMR·B A Johnson, R A Blevins

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Citations

Jul 29, 2008·The Journal of Biological Chemistry·Christine R ScharCynthia B Peterson
Dec 13, 2007·Journal of Molecular Recognition : JMR·Rebecca L Rich, David G Myszka
Jun 15, 2007·Protein Science : a Publication of the Protein Society·Aiwu Zhou
Sep 4, 2007·Protein Science : a Publication of the Protein Society·Magnus KjaergaardMichael Ploug
Jun 24, 2010·Protein Science : a Publication of the Protein Society·Tinoush MoulaeiAlexander Wlodawer
Dec 26, 2006·The Journal of Biological Chemistry·Xiangqun LiWuyuan Lu

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