Measuring transmembrane helix interaction strengths in lipid bilayers using steric trapping

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

Research output: Chapter in Book/Report/Conference proceedingChapter

15 Citations (Scopus)

Abstract

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.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
Pages37-56
Number of pages20
Volume1063
DOIs
Publication statusPublished - 2013
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume1063
ISSN (Print)10643745

Fingerprint

Lipid Bilayers
Streptavidin
Competitive Binding
Biotin
Membrane Proteins
Mutation

Keywords

  • Biotinylation
  • Glycophorin A dimer
  • Membrane protein folding
  • Monovalent streptavidin
  • Steric trap
  • Transmembrane helix interaction

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

Cite this

Hong, H., Chang, Y-C., & Bowie, J. U. (2013). Measuring transmembrane helix interaction strengths in lipid bilayers using steric trapping. In Methods in Molecular Biology (Vol. 1063, pp. 37-56). (Methods in Molecular Biology; Vol. 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. Vol. 1063 2013. p. 37-56 (Methods in Molecular Biology; Vol. 1063).

Research output: Chapter in Book/Report/Conference proceedingChapter

Hong, H, Chang, Y-C & Bowie, JU 2013, Measuring transmembrane helix interaction strengths in lipid bilayers using steric trapping. in Methods in Molecular Biology. vol. 1063, Methods in Molecular Biology, vol. 1063, pp. 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. In Methods in Molecular Biology. Vol. 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. Vol. 1063 2013. pp. 37-56 (Methods in Molecular Biology).
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