Quantitative super-resolution imaging reveals protein stoichiometry and nanoscale morphology of assembling HIV-gag virions

Julia Gunzenhäuser, Nicolas Olivier, Thomas Pengo, Suliana Manley

Research output: Contribution to journalArticlepeer-review

47 Scopus citations

Abstract

The HIV structural protein Gag assembles to form spherical particles of radius ∼70 nm. During the assembly process, the number of Gag proteins increases over several orders of magnitude from a few at nucleation to thousands at completion. The challenge in studying protein assembly lies in the fact that current methods such as standard fluorescence or electron microscopy techniques cannot access all stages of the assembly process in a cellular context. Here, we demonstrate an approach using super-resolution fluorescence imaging that permits quantitative morphological and molecular counting analysis over a wide range of protein cluster sizes. We applied this technique to the analysis of hundreds of HIV-Gag clusters at the cellular plasma membrane, thus elucidating how different fluorescent labels can change the assembly of virions.

Original languageEnglish (US)
Pages (from-to)4705-4710
Number of pages6
JournalNano letters
Volume12
Issue number9
DOIs
StatePublished - Sep 12 2012

Keywords

  • HIV-Gag
  • Super-resolution imaging
  • protein assembly
  • protein counting

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