Atomic force microscopy differentiates discrete size distributions between membrane protein containing and empty nanolipoprotein particles

Craig D. Blanchette, Jenny A. Cappuccio, Edward A. Kuhn, Brent W. Segelke, W. Henry Benner, Brett A. Chromy, Matthew A Coleman, Graham Bench, Paul D. Hoeprich, Todd A. Sulchek

Research output: Contribution to journalArticle

28 Scopus citations

Abstract

To better understand the incorporation of membrane proteins into discoidal nanolipoprotein particles (NLPs) we have used atomic force microscopy (AFM) to image and analyze NLPs assembled in the presence of bacteriorhodopsin (bR), lipoprotein E4 n-terminal 22k fragment scaffold and DMPC lipid. The self-assembly process produced two distinct NLP populations: those containing inserted bR (bR-NLPs) and those that did not (empty-NLPs). The bR-NLPs were distinguishable from empty-NLPs by an average increase in height of 1.0 nm as measured by AFM. Streptavidin binding to biotinylated bR confirmed that the original 1.0 nm height increase corresponds to br-NLP incorporation. AFM and ion mobility spectrometry (IMS) measurements suggest that NLP size did not vary around a single mean but instead there were several subpopulations, which were separated by discrete diameters. Interestingly, when bR was present during assembly the diameter distribution was shifted to larger particles and the larger particles had a greater likelihood of containing bR than smaller particles, suggesting that membrane proteins alter the mechanism of NLP assembly.

Original languageEnglish (US)
Pages (from-to)724-731
Number of pages8
JournalBiochimica et Biophysica Acta - Biomembranes
Volume1788
Issue number3
DOIs
StatePublished - Mar 2009
Externally publishedYes

Keywords

  • Apolipoprotein
  • Atomic force microscopy
  • Bacteriorhodopsin
  • Membrane protein
  • Nanodisc
  • Nanolipoprotein particle
  • NLP

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Biophysics

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  • Cite this

    Blanchette, C. D., Cappuccio, J. A., Kuhn, E. A., Segelke, B. W., Benner, W. H., Chromy, B. A., Coleman, M. A., Bench, G., Hoeprich, P. D., & Sulchek, T. A. (2009). Atomic force microscopy differentiates discrete size distributions between membrane protein containing and empty nanolipoprotein particles. Biochimica et Biophysica Acta - Biomembranes, 1788(3), 724-731. https://doi.org/10.1016/j.bbamem.2008.11.019