NDLI: Stabilizing the integrin alpha M inserted domain in alternative conformations with a range of engineered disulfide bonds.

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Stabilizing the integrin alpha M inserted domain in alternative conformations with a range of engineered disulfide bonds.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Xiao, Tsan Springer, Timothy A. Lu, Chafen Salas, Azucena Shimaoka, Motomu Takagi, Junichi
Description	Author Affiliation: Shimaoka M ( Center for Blood Research, Departments of Pathology, Pediatrics, and Anesthesia, Harvard Medical School, 200 Longwood Avenue, Boston, MA 02115, USA.);
Abstract	Conformational movement of the C-terminal alpha7 helix in the integrin inserted (I) domain, a major ligand-binding domain that adopts an alpha/beta Rossmann fold, has been proposed to allosterically regulate ligand-binding activity. Disulfide bonds were engineered here to reversibly lock the position of the alpha7 helix in one of two alternative conformations seen in crystal structures, termed open and closed. Our results show that pairs of residues with Cbeta atoms farther apart than optimal for disulfide bond stereochemistry can be successfully replaced by cysteine, suggesting that backbone movement accommodates disulfide formation. We also find more success with substituting partially exposed than buried residues. Disulfides stabilizing the open conformation resulted in constitutively active alphaMbeta2 heterodimers and isolated alphaM inserted domains, which were reverted to an inactive form by dithiothreitol reduction. By contrast, a disulfide stabilizing the closed conformation resulted in inactive alphaMbeta2 that was resistant to activation but became activatable after dithiothreitol treatment.
ISSN	00278424
e-ISSN	10916490
Journal	Proceedings of the National Academy of Sciences of the United States of America
Issue Number	26
Volume Number	99
Language	English
Publisher	National Academy of Sciences
Publisher Date	2002-12-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Macrophage-1 Antigen Chemistry Dimerization Disulfides Dithiothreitol Pharmacology Physiology Protein Conformation Protein Engineering Research Support, U.S. Gov't, P.H.S. Multidisciplinary
Content Type	Text
Resource Type	Article
Subject	Multidisciplinary

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