Purification and characterization of chaperonins 60 and 10 from Methylobacillus glycogenes.
Two proteins belonging to the group I chaperonin family were isolated from an obligate methanotroph, Methylobacillus glycogenes. The two proteins, one a GroEL homologue (cpn60: M. glycogenes 60 kDa chaperonin) and the other a GroES homologue (cpn10: M. glycogenes 10 kDa chaperonin), composed a heteropolymeric complex in the presence of ATP. Both proteins were purified from crude extracts of M. glycogenes by anion-exchange (DEAE-Toyopearl) and gel-filtration (Sephacryl S-400) chromatography. The native molecular weights of each chaperonin protein as determined by high-performance liquid chromatography (HPLC) gel-filtration were 820 000 for cpn60 and 65 000 for cpn10. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis revealed that the subunit molecular weights of cpn60 and cpn10 were 58 000 and 10 000, respectively. Both cpn60 and cpn10 possessed amino acid sequences which were highly homologous to other group I chaperonins. M. glycogenes cpn60 displayed an ATPase activity which was inhibited in the presence of cpn10. The chaperonins also displayed an ability to interact with and facilitate the refolding of Thermus malate dehydrogenase and yeast enolase in a manner similar to that of GroEL/ES. The similarities between the Escherichia coli GroE proteins are discussed.
Department of Biotechnology, Faculty of Engineering, Tottori University, Japan. kawata@bio.tottori-u.ac.jp
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Y
Kawata
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Doi
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H
Omoto
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Mizobata
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Nagai
D000251:Adenosine Triphosphatases / Q000378:metabolism; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000302:isolation & purification*; D018835:Chaperonin 10 / Q000737:chemistry / Q000302:isolation & purification*; D018834:Chaperonin 60 / Q000737:chemistry / Q000302:isolation & purification*; D002850:Chromatography, Gel; D002851:Chromatography, High Pressure Liquid; D002852:Chromatography, Ion Exchange; D004926:Escherichia coli / Q000737:chemistry; D005656:Fungal Proteins / Q000737:chemistry; D006088:Gram-Negative Aerobic Bacteria / Q000737:chemistry*; D006801:Humans; D008291:Malate Dehydrogenase / Q000737:chemistry; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010751:Phosphopyruvate Hydratase / Q000737:chemistry; D017510:Protein Folding; D012441:Saccharomyces cerevisiae / Q000737:chemistry; D016415:Sequence Alignment; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity
D001426:Bacterial Proteins; D018835:Chaperonin 10; D018834:Chaperonin 60; D005656:Fungal Proteins; D008291:Malate Dehydrogenase; D000251:Adenosine Triphosphatases; D010751:Phosphopyruvate Hydratase