NDLI: Functional Unfolding of α1-Antitrypsin Probed by Hydrogen-Deuterium Exchange Coupled with Mass Spectrometry* S⃞

Content Provider	PubMed Central
Author	Baek, Je-hyun Yang, Won Suk Lee, Cheolju Yu, Myeong-hee
Abstract	The native state of α1-antitrypsin (α1AT), a member of the serine protease inhibitor (serpin) family, is considered a kinetically trapped folding intermediate that converts to a more stable form upon complex formation with a target protease. Although previous structural and mutational studies of α1AT revealed the structural basis of the native strain and the kinetic trap, the mechanism of how the native molecule overcomes the kinetic barrier to reach the final stable conformation during complex formation remains unknown. We hypothesized that during complex formation, a substantial portion of the molecule undergoes unfolding, which we dubbed functional unfolding. Hydrogen-deuterium exchange coupled with ESI-MS was used to analyze this serpin in three forms: native, complexing, and complexed with bovine β-trypsin. Comparing the deuterium content at the corresponding regions of these three samples, we probed the unfolding of α1AT during complex formation. A substantial portion of the α1AT molecule unfolded transiently during complex formation, including not only the regions expected from previous structural studies, such as the reactive site loop, helix F, and the following loop, but also regions not predicted previously, such as helix A, strand 6 of β-sheet B, and the N terminus. Such unfolding of the native interactions may elevate the free energy level of the kinetically trapped native serpin sufficiently to cross the transition state during complex formation. In the current study, we provide evidence that protein unfolding has to accompany functional execution of the protein molecule.
Related Links	http://dx.doi.org/10.1074/mcp.m800365-mcp200
Ending Page	1081
Page Count	10
Starting Page	1072
File Format	PDF
ISSN	15359476
e-ISSN	15359484
Journal	Molecular & Cellular Proteomics : MCP
Issue Number	5
Volume Number	8
Language	English
Publisher	American Society for Biochemistry and Molecular Biology
Publisher Date	2009-05-01
Access Restriction	Open
Rights Holder	American Society for Biochemistry and Molecular Biology
Subject Keyword	Analytical Chemistry Biochemistry Molecular Biology Research in Higher Education
Content Type	Text
Resource Type	Article
Subject	Medicine Analytical Chemistry Molecular Biology Biochemistry

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