「BIOPHYSICS」に Atsushi Mukaiyama et al."A protocol for preparing nucleotide-free KaiC monomer"を掲載

2015 March 23 BPPB


Atsushi Mukaiyama, Masato Osako, Takaaki Hikima, Takao Kondo, Shuji Akiyama
”A protocol for preparing nucleotide-free KaiC monomer”

Intact KaiC hexamer reconstructed from nucleotide-free KaiC monomer stabilized in an optimized buffer system

KaiC, a core protein of the cyanobacterial circadian clock, self-assembles into a hexamer upon binding ATP. The instability of a nucleotide-free KaiC monomer has precluded its storage and further given rise to the difficulties of detailed analyses of the enzymatic properties of the KaiC hexamer. Here, we demonstrate the successful preparation of the nucleotide-free KaiC monomer in a phosphate buffer containing glutamic acid and arginine, and show that the intact KaiC hexamer is reconstructed from the prepared monomer upon addition of ATP. This protocol provides a methodological basis for further understanding of the structure and function of the KaiC hexamer.

BIOPHYSICS, Vol.11, pp. 79-84


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