Label-Free Single-Molecule Pulldown for the Detection of Released Cellular Protein Complexes

Guangzhong Ma, Pengfei Zhang, Xinyu Zhou, Zijian Wan, Shaopeng Wang

Research output: Contribution to journalArticlepeer-review

Abstract

Precise and sensitive detection of intracellular proteins and complexes is key to the understanding of signaling pathways and cell functions. Here, we present a label-free single-molecule pulldown (LFSMP) technique for the imaging of released cellular protein and protein complexes with single-molecule sensitivity and low sample consumption down to a few cells per mm2. LFSMP is based on plasmonic scattering imaging and thus can directly image the surface-captured molecules without labels and quantify the binding kinetics. In this paper, we demonstrate the detection principle for LFSMP, study the phosphorylation of protein complexes involved in a signaling pathway, and investigate how kinetic analysis can be used to improve the pulldown specificity. We wish our technique can contribute to uncovering the molecular mechanisms in cells with single-molecule resolution.

Original languageEnglish (US)
Pages (from-to)1272-1281
Number of pages10
JournalACS Central Science
Volume8
Issue number9
DOIs
StatePublished - Sep 28 2022
Externally publishedYes

ASJC Scopus subject areas

  • Chemistry(all)
  • Chemical Engineering(all)

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