Cloning, expression and purification of three Chaperonin 60 homologues.
The Chaperonin 60 (Cpn60) proteins have, in addition to their well-known functions of protein folding and protection, a range of intercellular signalling activities. As part of a study to investigate the biological activity of the Cpn60 proteins, particularly from pathogenic organisms, we have cloned and expressed three Cpn60 proteins from Homo sapiens, Helicobacter pylori and Chlamydia pneumoniae. The Cpn60 proteins were purified to apparent homogeneity using a combination of nickel column affinity chromatography and Reactive Red dye affinity columns. Insoluble protein was solubilised using 8 M urea and then re-folded on the nickel column by stepwise removal of the urea. The immunostimulant LPS was removed by addition of the antibiotic polymyxin B as part of the purification process.
Maguire
Maria
M
Cellular Microbiology Research Group, Eastman Dental Institute, University College London, 256 Grays Inn Road, WC1X 8LD, London, UK.
Coates
Anthony R M
AR
Henderson
Brian
B
D001483:Base Sequence; D018834:Chaperonin 60 / Q000235:genetics* / Q000302:isolation & purification*; D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D003001:Cloning, Molecular; D017931:DNA Primers; D004591:Electrophoresis, Polyacrylamide Gel; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification
D018834:Chaperonin 60; D017931:DNA Primers; D011994:Recombinant Proteins