[2P-653] Photo-control of Mitotic Kinesin Eg5 using Photoresponsive Protein (Aureochrome1 and Dronpa 145N).Department of Biosciences, Graduate School of Science and Engineering, Soka University, Hachioji, Tokyo, Japan.Soka university, Faculty of Science and Engineering, Department of science and Engineering for Sustainable Innovation.Besong Stanley Tabi, Islam MD Alrazi, Nobuyuki Nishibe and Shinsaku MarutaMitotic Kinesin Eg5 is a member of kinesin superfamily that is essential for Mitosis. Kinesin Eg5 is a motor protein that is homotetramer in structure which help during cross-links of anti-parallel microtubules in the mitotic spindle to maintain spindle bipolarity. Previously we have incorporated photochromic compounds of azobenzene derivatives into the functional site of Eg5 and controlled the Eg5 ATPase activity photoreversible. Aueochrome1 and Dronpa are photo responsive proteins and considered as a photochromic molecular device. It is known that the variants of aureochrome1, photozipper (PZ) and variant of Dronpa, Dronpa145N exhibit monomer-dimer conversion photoreversibly. In this study we tried to control the activity of kinesin Eg5 by introducing the variants of the photochromic protein into C-terminal of Eg5 motor domain. The designed fusion proteins of Eg5-PZ and Eg5-Dronpa145N were successfully expressed by E. coli expression system. Eg5-PZ showed spectral changes upon blue light irradiation and in the dark indicating photoisomerization. Monomer-dimer conversion of Eg5-PZ accompanied by photoisomerization was also observed by using size exclusion chromatography. However, ATPase activity of Eg5-PZ was not changed by blue light irradiation and in the dark. On the other hand, Eg5-Dronpa145N did not show spectral change upon 400 nm and 500 nm alternate light irradiations indicating that Eg5-Dronpa145N does not exhibit photoisomerization.

^〇STANLEY TABI BESONG ¹, Islam MD Alrazi¹, Nobuyuki Nishibe¹, Shinsaku Maruta¹ (Soka university Japan )

キーテクノロジー：HPLC,spectrophotometer

Kinesin Eg5、Aureochrome-1 、ATP ASSAY、Dronpa 145N、Isomerism