TITLE:
Effective cofactor complex purification using nanobeads
AUTHORS:
Makoto Umeda, Tatsuya Uebi, Naoya Maekawa, Yuka Masaike, Hiroshi Handa, Takeshi Imai
KEYWORDS:
Arginine; Nanobeads
JOURNAL NAME:
Journal of Biosciences and Medicines,
Vol.1 No.3,
December
12,
2013
ABSTRACT:
Drug target factor complex identification is necessary for
evidence based drug discovery. Previous study showed that using small chemical
immobilized magnetic
nanobeads the chemical target factors were effectively purified and identified. Here we succeeded to purify the
chemical target factor complex, so called cofactor(s). Arginine exhibits a variety of biological
activities through a complex and highly regulated set of pathways that remain
incompletely understood at both the whole-body and the cellular levels. The aim
of this study is to develop and validate effective purification system for arginine target
complex. New arginine
target protein (arginine interacting factor 4, AIF4) was purified and
identified. Using recombinant AIF4 protein and arginine-immobilized magnetic
nanobeads, AIF4 cofactor, AIF4-BP1, were purified. Interaction of AIF4 and
AIF4-BP1 was detected in arginine-dependent manner, suggesting arginine receptor complex formation. This nanobeads
technology is more than
30-fold efficient purification efficient than general purification technology.