title: A simple and rapid method for the purification of GroEL, an Escherichia coli homolog of the heat shock protein 60 family of molecular chaperonins.
abstract: GroEL, an Escherichia coli homolog of the heat shock protein 60 family of molecular chaperonins, has been implicated as a target of T cell-mediated immune responses in a broad spectrum of infections. In order to produce large quantities of native protein for raising and stimulating GroEL specific T cell lines, we have developed a simple and rapid two-step protocol for purifying native E. coli GroEL heat shock (or stress) protein which takes advantage of the inherent structural and functional properties of the protein. Based on a combination of gel exclusion chromatography, ATPase activity assay, isoelectric focusing, and circular dichroism analyses we conclude that our purification process yields native tetradecameric GroEL.
authors.affiliation: Procept, Inc., Cambridge, Massachusetts 02139.
authors.forename: S S
authors.initials: SS
authors.lastname: Khandekar
authors.forename: B M
authors.initials: BM
authors.lastname: Bettencourt
authors.forename: K C
authors.initials: KC
authors.lastname: Kelley
authors.forename: M A
authors.initials: MA
authors.lastname: Recny
mesh_terms: D000255:Adenosine Triphosphate / Q000378:metabolism; D001426:Bacterial Proteins / Q000302:isolation & purification*; D018834:Chaperonin 60; D002942:Circular Dichroism; D004926:Escherichia coli / Q000737:chemistry*; D006360:Heat-Shock Proteins / Q000302:isolation & purification*; D007525:Isoelectric Focusing; D008970:Molecular Weight; D011487:Protein Conformation; D013056:Spectrophotometry, Ultraviolet
chemical_list: D001426:Bacterial Proteins; D018834:Chaperonin 60; D006360:Heat-Shock Proteins; D000255:Adenosine Triphosphate