Sorting sub-150-nm liposomes of distinct sizes by DNA-brick-assisted centrifugation

Nat Chem. 2021 Apr;13(4):335-342. doi: 10.1038/s41557-021-00667-5. Epub 2021 Mar 30.

Abstract

In cells, myriad membrane-interacting proteins generate and maintain curved membrane domains with radii of curvature around or below 50 nm. To understand how such highly curved membranes modulate specific protein functions, and vice versa, it is imperative to use small liposomes with precisely defined attributes as model membranes. Here, we report a versatile and scalable sorting technique that uses cholesterol-modified DNA 'nanobricks' to differentiate hetero-sized liposomes by their buoyant densities. This method separates milligrams of liposomes, regardless of their origins and chemical compositions, into six to eight homogeneous populations with mean diameters of 30-130 nm. We show that these uniform, leak-resistant liposomes serve as ideal substrates to study, with an unprecedented resolution, how membrane curvature influences peripheral (ATG3) and integral (SNARE) membrane protein activities. Compared with conventional methods, our sorting technique represents a streamlined process to achieve superior liposome size uniformity, which benefits research in membrane biology and the development of liposomal drug-delivery systems.

Publication types

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

MeSH terms

  • Autophagy-Related Protein 7 / metabolism
  • Centrifugation / methods*
  • Cholesterol / analogs & derivatives
  • DNA / chemistry*
  • Liposomes / isolation & purification*
  • Liposomes / metabolism
  • Particle Size
  • SNARE Proteins / metabolism

Substances

  • Liposomes
  • SNARE Proteins
  • DNA
  • Cholesterol
  • Autophagy-Related Protein 7