title: Isolation and solubilization of gram-positive bacterial cell wall-associated proteins.
abstract: This chapter describes a simple, rapid and reproducible method to prepare bacterial cell wall extracts for two-dimensional gel electrophoresis (2DE). The extraction process uses mutanolysin, an N-acetylmuramidase, to gently solubilize cell wall-associated proteins from Gram-positive prokaryotes. The cells are first washed with buffer and resuspended in a solution containing mutanolysin. Following incubation at 37 degrees C, the sample is centrifuged and the supernatant containing the soluble cell wall-associated proteins is harvested. Following a brief precipitation step, the pellet is solubilized in sample buffer ready for isoelectric focusing and 2DE analysis.
authors: University of Wollongong, Wollongong, Australia.
authors: Jason N
authors: JN
authors: Cole
authors: Steven P
authors: SP
authors: Djordjevic
authors: Mark J
authors: MJ
authors: Walker
mesh_terms: D001426:Bacterial Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D002473:Cell Wall / Q000737:chemistry*; D015180:Electrophoresis, Gel, Two-Dimensional; D012995:Solubility; D013297:Streptococcus pyogenes / Q000378:metabolism*
chemical_list: D001426:Bacterial Proteins