Protein Crystallization of Two Recombinant Lpt Proteins

Methods Mol Biol. 2022:2548:249-263. doi: 10.1007/978-1-0716-2581-1_15.

Abstract

The prerequisite for 3D structure determination of macromolecules via X-ray crystallography is well-ordered, diffracting crystals. Here, we report the recombinant production, biophysical/biochemical protein sample characterization, and vapor diffusion sitting drop crystallization protocols for two lipopolysaccharide transport proteins: LptH from Pseudomonas aeruginosa (Pa-LptH) and an inactive LptC mutant (G153R) from Escherichia coli (EcLptC24-191G153R).

Keywords: E. coli; Lipopolysaccharide transport protein; Outer membrane biogenesis; Protein crystallization; Vapor diffusion.

MeSH terms

  • Carrier Proteins / metabolism
  • Crystallization
  • Crystallography, X-Ray
  • Escherichia coli / genetics
  • Escherichia coli / metabolism
  • Escherichia coli Proteins* / metabolism
  • Lipopolysaccharides* / chemistry
  • Membrane Proteins / metabolism
  • Recombinant Proteins / genetics
  • Recombinant Proteins / metabolism

Substances

  • Carrier Proteins
  • Escherichia coli Proteins
  • Lipopolysaccharides
  • LptC protein, E coli
  • Membrane Proteins
  • Recombinant Proteins