Microfluidic measurement of the size and shape of lipid-anchored proteins

Biophys J. 2024 Oct 1;123(19):3478-3489. doi: 10.1016/j.bpj.2024.08.026. Epub 2024 Sep 2.

Abstract

The surface of a cell is crowded with membrane proteins. The size, shape, density, and mobility of extracellular surface proteins mediate cell surface accessibility to external molecules, viral particles, and other cells. However, predicting these qualities is not always straightforward, even when protein structures are known. We previously developed an experimental method for measuring flow-driven lateral transport of neutravidin bound to biotinylated lipids in supported lipid bilayers. Here, we use this method to detect hydrodynamic force applied to a series of lipid-anchored proteins with increasing size. We find that the measured force reflects both protein size and shape, making it possible to distinguish these features of intact, folded proteins in their undisturbed orientation and proximity to the lipid membrane. In addition, our results demonstrate that individual proteins are transported large distances by flow forces on the order of femtoNewtons, similar in magnitude to the shear forces resulting from blood circulation or from the swimming motion of microorganisms. Similar protein transport across living cells by hydrodynamic force may contribute to biological flow sensing.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, N.I.H., Extramural

MeSH terms

  • Avidin / chemistry
  • Avidin / metabolism
  • Hydrodynamics*
  • Lipid Bilayers / chemistry
  • Lipid Bilayers / metabolism
  • Membrane Proteins / chemistry
  • Membrane Proteins / metabolism

Substances

  • Lipid Bilayers
  • Membrane Proteins
  • Avidin