Molecular cloning of groESL locus, and purification and characterization of chaperonins, GroEL and GroES, from Bacillus brevis
Overview of Tokunaga M et al.
| Authors | Tokunaga M Shiraishi Y Odachi M Mizukami M Tokunaga H Philo JS Arakawa T Ishibashi M Tanaka R Takagi H |
|---|---|
| Affiliation | Laboratory of Applied and Molecular Microbiology Faculty of Agriculture Kagoshima University Korimoto Japan. tokunaga@chem.agri.kagoshima-u.ac.jp |
| Journal | Biosci Biotechnol Biochem |
| Year | 2001 |
Abstract
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.