Analysis of Bacterial Cell Surface Chemical Composition Using Cryogenic X-Ray Photoelectron Spectroscopy.
This chapter describes a method for measuring the average surface chemical composition with respect to lipids, polysaccharides, and peptides (protein + peptidoglycan) for the outer part of the bacterial cell wall. Bacterial cultures grown over night are washed with a buffer or saline at controlled pH. The analysis is done on fast-frozen bacterial cell pellets obtained after centrifugation, and the analysis requires access to X-ray photoelectron spectroscopy instrumentation that can perform analyses at cryogenic temperatures (for example using liquid nitrogen). The method can be used to monitor changes in the cell wall composition following environmental stimuli or genetic mutations. The data obtained originate from the outermost part of the cell wall. Thus, it is expected that for gram-negative bacteria only the outer membrane and part of the periplasmic peptidoglycan layer is probed during analysis, and for gram-positive bacteria only the top nanometers of the peptidoglycan layer of the cell wall is monitored.
Department of Chemistry, Umeå University, Linnaeus väg 10, 901 87, Umeå, Sweden. madeleine.ramstedt@chem.umu.se.
Madeleine
M
Ramstedt
Department of Chemistry, Umeå University, Linnaeus väg 10, 901 87, Umeå, Sweden.
Andrey
A
Shchukarev
D001426:Bacterial Proteins / Q000032:analysis; D002462:Cell Membrane / Q000737:chemistry; D002473:Cell Wall / Q000737:chemistry*; D006090:Gram-Negative Bacteria / Q000737:chemistry / Q000254:growth & development; D006094:Gram-Positive Bacteria / Q000737:chemistry / Q000254:growth & development; D008055:Lipids / Q000032:analysis*; D010455:Peptides / Q000032:analysis*; D056951:Photoelectron Spectroscopy; D011135:Polysaccharides, Bacterial / Q000032:analysis*
D001426:Bacterial Proteins; D008055:Lipids; D010455:Peptides; D011135:Polysaccharides, Bacterial
Bacterial cells; C 1s spectra; Cell wall composition; X-ray photoelectron spectroscopy