Use of surface‐enhanced laser desorption/ionization ‐time of flight to explore bacterial proteomes

Abstract

Surface-enhanced laser desorption/ionization (SELDI)-time of flight is a recent technology that allows proteomic analysis with limited material requirements. This characteristic makes it a valuable technique for microbiologists handling problematic samples, such as low cell number cultures. We compared three simple procedures for protein extraction from bacteria for compatibility with the ProteinChip Array; we also determined the amount of protein required for each analysis. The protocol for the SELDI analysis was evaluated by generating protein expression profiles of a Streptococcus pneumoniae strain grown in different conditions and those of different strains of the same species. The protocol also was successfully applied to a wide range of Gram positive and negative bacteria. The results of this study suggest the appropriateness of this technology for microorganism protein profiling as complementary or alternative to two-dimensional gel electrophoresis. Abbreviations: BHI, brain heart infusion; EAM, energy absorbing molecule; GC-TR, GramCracker and Tri-Reagent-LS method; SAX, strong anion exchange; SPA, sinapinic acid; WCX, weak cation exchange

Metamorphosis of Borrelia burgdorferi organisms – RNA, lipid and protein composition in context with the spirochetes' shape

“…With this accurate method according to mass, reproducibly and high output, the differential protein expression as well as up-and down-regulated proteins can be detected according to mass in a semiquantitative fashion. It has been recommended as an appropriate technology for the study of bacterial proteomics [44]. In our study, the profile of the proteins detected with this technique ranged between 6 -16 kDa.…”

Differences in protein profiles of the isolates of Entamoeba histolytica and E. dispar by surface-enhanced laser desorption ionization time-of-flight mass spectrometry (SELDI-TOF MS) ProteinChip assays

“…Using the different array surfaces, a complete picture of each strain of Entameoba may be drawn, and thus a better set of protein fingerprint profiles for each strain would be provided. Like the successful application of this technology to bacterial proteomes (Barzaghi et al 2004), the results show the usefulness of ProteinChip technology for studying the proteomics of parasitic protozoa as well, alone or in combination with other technologies.…”

Proteomics in Food Science

“…MALDI-TOF MS has been reported as the most widely used mass spectral method for bacterial identification and differentiation at the genus, species, and, in some cases, strain level due to its speed and reduced cost when compared with biochemical and molecular techniques and when coupled with other methodologies, such as SELDI-TOF MS, which is a modified version of MALDI-TOF MS that correctly identifies very closely related bacterial species (Barzaghi et al, 2004;Kiehntopf et al, 2011;Lundquist et al, 2005). Over the past 16 years, there have been several reports targeted mainly at clinical diagnostic microbiology, biodefence, and environmental research that have demonstrated the efficacy of MALDI-TOF MS as a tool for bacterial identification (Croxatto et al, 2011;Fenselau, 2008a, 2008b;Giebel et al, 2010;Freiwald and Sauer, 2009;Sauer et al, 2008;Welker and Moore, 2011), and many books on this field have recently appeared (Wilkins et al, 2006;Shah and Gharbia, 2010).…”