Measuring transmembrane helix interaction strengths in lipid bilayers using steric trapping

Heedeok Hong, Yu-Chu Chang, James U. Bowie

研究成果: 書貢獻/報告類型章節

15 引文 (Scopus)

摘要

We have developed a method to measure strong transmembrane (TM) helix interaction affinities in lipid bilayers that are difficult to measure by traditional dilution methods. The method, called steric trapping, couples dissociation of biotinylated TM helices to a competitive binding by monovalent streptavidin (mSA), so that dissociation is driven by the affinity of mSA for biotin and mSA concentration. By adjusting the binding affinity of mSA through mutation, the method can obtain dissociation constants of TM helix dimers (Kd,dimer) over a range of six orders of magnitudes. The K d,dimer limit of measurable target interaction is extended 3-4 orders of magnitude lower than possible by dilution methods. Thus, steric trapping opens up new opportunities to study the folding and assembly of α-helical membrane proteins in lipid bilayer environments. Here we provide detailed methods for applying steric trapping to a TM helix dimer.
原文英語
主出版物標題Methods in Molecular Biology
頁面37-56
頁數20
1063
DOIs
出版狀態已發佈 - 2013
對外發佈Yes

出版系列

名字Methods in Molecular Biology
1063
ISSN(列印)10643745

指紋

Lipid Bilayers
Streptavidin
Competitive Binding
Biotin
Membrane Proteins
Mutation

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

引用此文

Hong, H., Chang, Y-C., & Bowie, J. U. (2013). Measuring transmembrane helix interaction strengths in lipid bilayers using steric trapping. 於 Methods in Molecular Biology (卷 1063, 頁 37-56). (Methods in Molecular Biology; 卷 1063). https://doi.org/10.1007/978-1-62703-583-5_3

Measuring transmembrane helix interaction strengths in lipid bilayers using steric trapping. / Hong, Heedeok; Chang, Yu-Chu; Bowie, James U.

Methods in Molecular Biology. 卷 1063 2013. p. 37-56 (Methods in Molecular Biology; 卷 1063).

研究成果: 書貢獻/報告類型章節

Hong, H, Chang, Y-C & Bowie, JU 2013, Measuring transmembrane helix interaction strengths in lipid bilayers using steric trapping. 於 Methods in Molecular Biology. 卷 1063, Methods in Molecular Biology, 卷 1063, 頁 37-56. https://doi.org/10.1007/978-1-62703-583-5_3
Hong H, Chang Y-C, Bowie JU. Measuring transmembrane helix interaction strengths in lipid bilayers using steric trapping. 於 Methods in Molecular Biology. 卷 1063. 2013. p. 37-56. (Methods in Molecular Biology). https://doi.org/10.1007/978-1-62703-583-5_3
Hong, Heedeok ; Chang, Yu-Chu ; Bowie, James U. / Measuring transmembrane helix interaction strengths in lipid bilayers using steric trapping. Methods in Molecular Biology. 卷 1063 2013. 頁 37-56 (Methods in Molecular Biology).
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