title: Enhanced proteome profiling by inhibiting proteolysis with small heat shock proteins.
abstract: Proteolytic degradation is one of the critical problems in two-dimensional electrophoresis (2-DE). Here we report that small heat shock proteins (sHsps), including IbpA(Ec) and IbpB(Ec) from Escherichia coli and Hsp26(Sc) from Saccharomyces cerevisiae, are able to protect proteins in vitro from proteolytic degradation. Addition of sHsps during 2-DE of human serum or whole cell extracts of E. coli, Mannheimia succinciproducens, Arabidopsis thaliana, and human kidney cells allowed detection of up to 50% more protein spots than those obtainable with currently available protease inhibitors. Therefore, the use of sHsps during 2-DE significantly improves proteome profiling by generally enabling the detection of many more protein spots that could not be seen previously.
authors.affiliation: Metabolic and Biomolecular Engineering National Research Laboratory, Department of Chemical & Biomolecular Engineering, BioProcess Engineering Research Center, Daejeon, Republic of Korea.
authors.forename: Mee-Jung
authors.initials: MJ
authors.lastname: Han
authors.forename: Jeong Wook
authors.initials: JW
authors.lastname: Lee
authors.forename: Sang Yup
authors.initials: SY
authors.lastname: Lee
mesh_terms: D000818:Animals; D017360:Arabidopsis / Q000378:metabolism; D002460:Cell Line; D015180:Electrophoresis, Gel, Two-Dimensional; D004926:Escherichia coli / Q000378:metabolism; D006360:Heat-Shock Proteins / Q000737:chemistry*; D006801:Humans; D031121:Mannheimia / Q000378:metabolism; D011506:Proteins / Q000737:chemistry; D020543:Proteome; D040901:Proteomics / Q000379:methods*; D012441:Saccharomyces cerevisiae / Q000378:metabolism
chemical_list: D006360:Heat-Shock Proteins; D011506:Proteins; D020543:Proteome