title: Periplasmic localization of a GroES homologue in Escherichia coli transformed with groESx cloned from Legionella-like endosymbionts in Amoeba proteus.
abstract: Escherichia coli MC4100 transformed with a groE homologous operon cloned from X-bacteria accumulated large amounts of the gene product when cultured at 30 or 37 degrees C. Heat shock for 10-30 min at 42 degrees C or ethanol (5%) shock for 2 h increased GroESx levels to about twice that in E. coli grown at 30 degrees C. The subcellular localization of GroESx in transformed E. coli was determined by several subcellular fractionation methods, by the analysis of extracted proteins in SDS polyacrylamide gels and by assays of marker enzymes. The GroESx protein was detected in both the periplasmic and cytoplasmic extracts and a large amount of the protein was accumulated in the periplasm. The GroEL protein and recombinant beta-galactosidase were exclusively localized in the cytoplasmic fraction, eliminating the possibility that periplasmic GroESx might be due to simple overproduction. N-terminal amino acid sequencing confirmed that the protein resolved on a 2-D gel was GroESx. This work represents the first report of the periplasmic location of GroES homologues in E. coli.
authors.affiliation: School of Biological Sciences, Seoul National University, South Korea.
authors.forename: J E
authors.initials: JE
authors.lastname: Lee
authors.forename: T I
authors.initials: TI
authors.lastname: Ahn
mesh_terms: D000656:Amoeba / Q000235:genetics*; D000818:Animals; D018835:Chaperonin 10 / Q000032:analysis* / Q000235:genetics / Q000502:physiology; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D006358:Hot Temperature; D007875:Legionella / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis; D013559:Symbiosis; D014169:Transformation, Bacterial
chemical_list: D018835:Chaperonin 10; D011994:Recombinant Proteins