Observing lysozyme's closing and opening motions by high-resolution single-molecule enzymology

ACS Chemical Biology
Maxim V AkhterovPhilip G Collins

Abstract

Single-molecule techniques can monitor the kinetics of transitions between enzyme open and closed conformations, but such methods usually lack the resolution to observe the underlying transition pathway or intermediate conformational dynamics. We have used a 1 MHz bandwidth carbon nanotube transistor to electronically monitor single molecules of the enzyme T4 lysozyme as it processes substrate. An experimental resolution of 2 μs allowed the direct recording of lysozyme's opening and closing transitions. Unexpectedly, both motions required 37 μs, on average. The distribution of transition durations was also independent of the enzyme's state: either catalytic or nonproductive. The observation of smooth, continuous transitions suggests a concerted mechanism for glycoside hydrolysis with lysozyme's two domains closing upon the polysaccharide substrate in its active site. We distinguish these smooth motions from a nonconcerted mechanism, observed in approximately 10% of lysozyme openings and closings, in which the enzyme pauses for an additional 40-140 μs in an intermediate, partially closed conformation. During intermediate forming events, the number of rate-limiting steps observed increases to four, consistent with four steps requ...Continue Reading

References

Feb 10, 1989·Science·M Matsumura, B W Matthews
Dec 6, 1994·Proceedings of the National Academy of Sciences of the United States of America·K SvobodaS M Block
Jan 1, 1995·Cold Spring Harbor Symposia on Quantitative Biology·M J Schnitzer, S M Block
Jul 24, 1997·Nature·M J Schnitzer, S M Block
Dec 4, 1998·Science·H P LuX S Xie
Feb 3, 1999·Proceedings of the National Academy of Sciences of the United States of America·T HaS Weiss
Jul 18, 2001·Journal of the American Chemical Society·R J ChenH Dai
Sep 15, 2004·Proceedings of the National Academy of Sciences of the United States of America·Fernando PatolskyCharles M Lieber
Oct 2, 2004·Science·Saveez SaffarianGregory Goldberg
Dec 20, 2005·Accounts of Chemical Research·Wei MinX Sunney Xie
Mar 16, 2006·Proceedings of the National Academy of Sciences of the United States of America·Samy O MerouehShahriar Mobashery
Dec 14, 2007·Nature·Katherine Henzler-Wildman, Dorothee Kern
May 31, 2008·Nature Methods·Rahul RoyTaekjip Ha
Jul 26, 2008·Nature Nanotechnology·Xinjian ZhouPaul L McEuen
Jul 9, 2009·Proceedings of the National Academy of Sciences of the United States of America·Hoi Sung ChungWilliam A Eaton
Apr 8, 2010·The Journal of Physical Chemistry. B·Yuanmin Wang, H Peter Lu
Jul 21, 2010·Biophysical Journal·Daniel L FloydAntoine M van Oijen
Nov 16, 2010·Nature Nanotechnology·Shuo HuangStuart Lindsay
Aug 3, 2011·Nano Letters·Sebastian SorgenfreiKenneth L Shepard
Jan 14, 2012·Journal of the American Chemical Society·Yongki ChoiGregory A Weiss
Jan 24, 2012·Science·Yongki ChoiPhilip G Collins
Feb 23, 2012·Scientific Reports·Makusu TsutsuiTomoji Kawai
Apr 14, 2012·Science·Morten Ø JensenDavid E Shaw
Oct 16, 2012·Biophysical Journal·Robel B Yirdaw, Hassane S McHaourab
Nov 8, 2012·Nature Nanotechnology·R Dean Astumian
Oct 25, 2013·Nature·Hoi Sung Chung, William A Eaton
Apr 8, 2014·Nature Nanotechnology·Yanan ZhaoStuart Lindsay
Apr 18, 2014·Nature·Hesam N MotlaghVincent J Hilser
Apr 23, 2014·Proceedings of the National Academy of Sciences of the United States of America·Daniel-Adriano SilvaXuhui Huang

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Citations

Mar 7, 2017·Chemical Communications : Chem Comm·Seungyong YouYongki Choi
Oct 27, 2017·Proceedings of the National Academy of Sciences of the United States of America·Ziad Ganim, Matthias Rief
Dec 6, 2017·Physical Biology·Bozhi TianYuanwen Jiang
Apr 15, 2018·Journal of the American Society for Mass Spectrometry·Jieutonne J ArcherRobert J Levis
Mar 8, 2020·Nature Communications·Hugo SanabriaClaus A M Seidel
May 27, 2021·The Journal of Physical Chemistry. B·Sung Oh WooYongki Choi

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