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Interdomain flip-flop motion visualized in flavocytochrome cellobiose dehydrogenase using high-speed atomic force
Hirofumi Harada1, Akira Onoda1, Takayuki Uchihashi2
1Department of Applied Chemistry , Graduate School of Engineering , Osaka University , 2-1 Yamadaoka , Suita , Osaka 565-0871 , Japan . Email: onoda@chem.eng.osaka-u.ac.jp ;
Cellobiose dehydrogenase (CDH) undergoes dynamic domain motion during catalysis. Enzyme activity is triggered by cellobiose, enabling efficient electron transfer for external acceptors.
Area of Science:
- Biochemistry
- Enzymology
- Biophysics
Background:
- Cellobiose dehydrogenase (CDH) is a dual-domain flavocytochrome with dehydrogenase (DH) and cytochrome (CYT) domains.
- Understanding enzyme dynamics is crucial for elucidating catalytic mechanisms.
Purpose of the Study:
- To visualize the dynamic domain motion of Phanerochaete chrysosporium CDH (PcCDH) during catalysis.
- To investigate the role of cellobiose in triggering enzyme domain movement.
Main Methods:
- High-speed atomic force microscopy (HS-AFM) was employed to observe PcCDH.
- PcCDH was immobilized on a heme-coated gold surface to fix CYT domain orientation.
Main Results:
- PcCDH domains were immobile in the absence of cellobiose.
- Cellobiose addition induced interdomain flip-flop motion, including association and dissociation.
Conclusions:
- Dynamic domain motion in CDH is essential for catalysis.
- This motion facilitates efficient electron transfer to external electron acceptors.
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