GroEL chaperone binding to beetle luciferases and the implications for refolding when co-expressed |
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Authors: | Venkatesh Balan Arifuzzaman Mohammad Mori Hirotada Taguchi Takahisa Ohmiya Yoshihiro |
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Institution: | Cell Dynamics Research Group, National Institute of Advanced Industrial Science and Technology, Ikeda, Osaka 563-8577, Japan. venkatesh_balan@hotmail.com |
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Abstract: | The folding of many proteins including luciferase in vivo requires the assistance of molecular chaperone proteins. To understand how a chaperone targets luciferase, we took three luciferases that give different bioluminescence with the same luciferin substrate and with differences in homology. The three luciferase genes, firefly luciferase (FF-Luc) (from Pyrocoelia miyako), and red (RE-Luc) and green (GR-Luc) bioluminescence-emitting luciferases (from Phrixothrix railroad-worms), were expressed in Escherichia coli to produce fusion proteins with predicted molecular masses. Subsequently, we observed that DnaK and GroEL were co-purified along with recombinant luciferase. Although the amount of co-purified DnaK was almost the same compared to FF-Luc, GroEL was 25 and 32 times higher in GR-Luc and RE-Luc respectively. Furthermore, co-expression of GroEL/GroES along with luciferase substantially refolded RE-Luc and GR-Luc compared to FF-Luc. |
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