Molecular Cloning of groESL Locus, and Purification and Characterization of Chaperonins, GroEL and GroES, from Bacillus brevis | Zendy

Masao Tokunaga | Zendy; Yoichi Shiraishi | Zendy; Masatake ODACHI | Zendy; Makoto Mizukami | Zendy; Hiroko Tokunaga | Zendy; John S. Philo | Zendy; Tsutomu Arakawa | Zendy; Matsujiro Ishibashi | Zendy; Ryoichi Tanaka | Zendy; Hiroaki Takagi | Zendy

Open Access

Molecular Cloning of groESL Locus, and Purification and Characterization of Chaperonins, GroEL and GroES, from Bacillus brevis

Author(s) -

Masao Tokunaga,

Yoichi Shiraishi,

Masatake ODACHI,

Makoto Mizukami,

Hiroko Tokunaga,

John S. Philo,

Tsutomu Arakawa,

Matsujiro Ishibashi,

Ryoichi Tanaka,

Hiroaki Takagi

Publication year - 2001

Publication title -

bioscience biotechnology and biochemistry

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 0.509

H-Index - 116

eISSN - 1347-6947

pISSN - 0916-8451

DOI - 10.1271/bbb.65.1379

Subject(s) - groel , groes , chaperonin , biology , microbiology and biotechnology , bacillus subtilis , biochemistry , protein folding , escherichia coli , bacteria , genetics , gene

The groESL locus of a protein-hypersecreting bacterium, Bacillus brevis, was cloned by PCR using primers designed based on the DNA sequence of a B. subtilis homolog. GroEL protein was purified to apparent homogeneity and its ATPase activity was characterized: it hydrolyzed ATP, CTP, and TTP in this order of reaction rate, and its specific activity for ATP was 0.1 micromole/min/mg protein. Purified GroEL forms a tetradecamer. GroEL was estimated to contain 22% alpha-helix, 24% beta-sheet, and 19% turn structures, by CD measurement. GroES protein was also highly purified to examine its chaperonin activity. GroEL protected from thermal inactivation of and showed refolding-promoting activity for malate dehydrogenase, strictly depending on the presence of ATP and GroES.

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