Structure retention of silica gel-encapsulated bacteriorhodopsin in purple membrane and in lipid nanodiscs

Colloids Surf B Biointerfaces. 2020 Feb;186:110680. doi: 10.1016/j.colsurfb.2019.110680. Epub 2019 Nov 27.

Abstract

The integral membrane protein, bacteriorhodopsin (BR) was encapsulated in sol-gel derived porous silica gel monoliths in native purple membrane (BR-PM) and synthetic lipid nanodisc (BR nanodisc) environments. BR nanodiscs were synthesized by solubilizing purple membrane in discoidal phospholipid bilayer stabilized by amphipathic Styrene-Maleic Acid (SMA) copolymer. UV-vis absorbance spectroscopy and dynamic-light scattering indicated the formation of BR monomers solubilized in lipid nanodiscs 10.2 ± 0.7 nm in average diameter. Fluorescence and absorbance spectroscopic techniques were utilized to probe conformational, environmental, and rotational changes associated with the tryptophan residues and the covalently-bound retinal moiety of BR upon entrapment in the silica matrix. We show that the immobilized BR in both membrane environments retained its bound retinal cofactor and the ability of the cofactor to undergo conformational changes upon light illumination necessary for BR's activity as a proton transporter. For purple membrane fragments, the results indicated that the local pH in the pores around BR after encapsulation was important for its stability at temperatures higher than 50 °C. Under the same buffering conditions, retinal was released from silica-encapsulated BR-PM and BR nanodiscs beginning at 80 °C (without a conformational change) and 50 °C (with a conformational change), respectively, reflecting differences in protein-protein (trimeric vs. monomeric) and protein-lipid interactions.

Keywords: Encapsulation; Hybrid biomaterial; Integral membrane protein; Lipid bilayer; Lipid nanodisc; SMA; Sol-Gel.

MeSH terms

  • Bacteriorhodopsins / chemistry*
  • Gels / chemistry
  • Lipids / chemistry*
  • Nanostructures / chemistry*
  • Particle Size
  • Purple Membrane / chemistry*
  • Silicon Dioxide / chemistry*
  • Surface Properties

Substances

  • Gels
  • Lipids
  • Bacteriorhodopsins
  • Silicon Dioxide