Controlled unfolding and refolding of a single sodium-proton antiporter using atomic force microscopy

Journal of Molecular Biology
Alexej KedrovDaniel J Muller

Abstract

Single-molecule force-spectroscopy was employed to unfold and refold single sodium-proton antiporters (NhaA) of Escherichia coli from membrane patches. Although transmembrane alpha-helices and extracellular polypeptide loops exhibited sufficient stability to individually establish potential barriers against unfolding, two helices predominantly unfolded pairwise, thereby acting as one structural unit. Many of the potential barriers were detected unfolding NhaA either from the C-terminal or the N-terminal end. It was found that some molecular interactions stabilizing secondary structural elements were directional, while others were not. Additionally, some interactions appeared to occur between the secondary structural elements. After unfolding ten of the 12 helices, the extracted polypeptide was allowed to refold back into the membrane. After five seconds, the refolded polypeptide established all secondary structure elements of the native protein. One helical pair showed a characteristic spring like "snap in" into its folded conformation, while the refolding process of other helices was not detected in particular. Additionally, individual helices required characteristic periods of time to fold. Correlating these results with the ...Continue Reading

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Citations

Mar 18, 2006·Naunyn-Schmiedeberg's Archives of Pharmacology·Alexej Kedrov, Daniel J Müller
May 15, 2007·Applied Microbiology and Biotechnology·Ana J García-Sáez, Petra Schwille
Dec 6, 2006·Biochemistry·Robert Renthal
Nov 3, 2010·Biochemistry·Shiho KawamuraPaul S-H Park
Feb 13, 2008·Langmuir : the ACS Journal of Surfaces and Colloids·K Tanuj SapraDaniel J Muller
Oct 12, 2011·Proceedings of the National Academy of Sciences of the United States of America·Lin GeDaniel J Muller
Oct 2, 2013·Proceedings of the National Academy of Sciences of the United States of America·Christian A BippesDaniel J Müller
May 8, 2009·The Journal of Biological Chemistry·Christian A BippesDimitrios Fotiadis
Mar 7, 2008·Pharmacological Reviews·Daniel J MüllerKrzysztof Palczewski
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Feb 22, 2007·Annual Review of Biophysics and Biomolecular Structure·Alexej KedrovDaniel J Müller
Aug 16, 2006·Proceedings of the National Academy of Sciences of the United States of America·Hendrik DietzMatthias Rief
Dec 18, 2007·Journal of Molecular Biology·Alexej KedrovDaniel J Muller
Jun 18, 2005·EMBO Reports·Alexej KedrovDaniel J Muller
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Sep 24, 2008·Nanotechnology·Jens StruckmeierDaniel J Müller

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