Functional unfolding of alpha1-antitrypsin probed by hydrogen-deuterium exchange coupled with mass spectrometry.

Molecular & Cellular Proteomics : MCP
Je-Hyun BaekMyeong-Hee Yu

Abstract

The native state of alpha(1)-antitrypsin (alpha(1)AT), 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 alpha(1)AT 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 beta-trypsin. Comparing the deuterium content at the corresponding regions of these three samples, we probed the unfolding of alpha(1)AT during complex formation. A substantial portion of the alpha(1)AT 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 n...Continue Reading

References

Jan 16, 1992·Nature·J MottonenE J Goldsmith
Jul 6, 1971·Biochemistry·N C RobinsonK A Walsh
Jul 1, 1995·Proteins·H T Wright, J N Scarsdale
Feb 1, 1995·Nature Structural Biology·P E Stein, R W Carrell
Dec 1, 1995·Trends in Biotechnology·R A EnghA J Schulze
Jun 1, 1996·Nature Structural Biology·K N LeeM H Yu
Feb 1, 1996·Current Opinion in Structural Biology·S W EnglanderL Mayne
Feb 1, 1997·Journal of Mass Spectrometry : JMS·D L SmithZ Zhang
May 1, 1997·Nature Structural Biology·D A Lawrence
Apr 10, 1999·The Journal of Biological Chemistry·H ImM H Yu
Apr 29, 1999·Proceedings of the National Academy of Sciences of the United States of America·E Stratikos, P G Gettins
Aug 10, 1999·Journal of the American Society for Mass Spectrometry·K A ResingN G Ahn
Jul 8, 2000·Proceedings of the National Academy of Sciences of the United States of America·C LeeM H Yu
Sep 7, 2000·Protein Science : a Publication of the Protein Society·H ImM H Yu
Oct 12, 2000·The Journal of Biological Chemistry·K M O'Malley, B S Cooperman
Feb 7, 2001·Proceedings of the National Academy of Sciences of the United States of America·A N HoofnagleN G Ahn
Jun 23, 2001·Protein Science : a Publication of the Protein Society·C LeeM H Yu
Jan 12, 2002·The Journal of Biological Chemistry·Jong-Shik Shin, Myeong-Hee Yu
Sep 24, 2002·The Journal of Biological Chemistry·Hana ImMyeong-Hee Yu
May 25, 2005·Biochemistry·Douglas V LaurentsMarta Bruix
Jun 9, 2007·EMBO Reports·Qingwei ZhangJames C Whisstock
Jun 27, 2007·Proceedings of the National Academy of Sciences of the United States of America·Paul SchandaBernhard Brutscher
Apr 23, 2008·Current Protocols in Protein Science·Jeffrey G MandellElizabeth A Komives
Sep 17, 2008·The Journal of Biological Chemistry·Yuko TsutsuiPatrick L Wintrode

❮ Previous
Next ❯

Citations

Jul 25, 2009·Biochemistry·Tanusree SenguptaPatrick L Wintrode
Oct 18, 2014·Nature Chemical Biology·Anne GershensonSheena E Radford
Nov 3, 2012·Biochimica Et Biophysica Acta·Sheila S Jaswal
Oct 15, 2014·Proceedings of the National Academy of Sciences of the United States of America·Giorgia CazzolliPatrick L Wintrode
May 5, 2012·Protein Expression and Purification·Ansgar Brock
Jul 23, 2014·Angewandte Chemie·Morten B TrelleThomas J D Jørgensen
Jul 19, 2011·International Journal of Mass Spectrometry·Anindya Sarkar, Patrick L Wintrode
Sep 3, 2014·PloS One·Victoria A HughesPatrick L Wintrode

❮ Previous
Next ❯

Related Concepts

Related Feeds

ASBMB Publications

The American Society for Biochemistry and Molecular Biology (ASBMB) includes the Journal of Biological Chemistry, Molecular & Cellular Proteomics, and the Journal of Lipid Research. Discover the latest research from ASBMB here.