Interdomain flip-flop motion visualized in flavocytochrome cellobiose dehydrogenase using high-speed atomic force microscopy during catalysis

Hirofumi Harada; Akira Onoda; Takayuki Uchihashi; Hiroki Watanabe; Naoki Sunagawa; Masahiro Samejima; Kiyohiko Igarashi; Takashi Hayashi

doi:10.1039/c7sc01672g

Interdomain flip-flop motion visualized in flavocytochrome cellobiose dehydrogenase using high-speed atomic force microscopy during catalysis

Hirofumi Harada
, Akira Onoda
, Takayuki Uchihashi
, Hiroki Watanabe
, Naoki Sunagawa
, Masahiro Samejima
, Kiyohiko Igarashi
, Takashi Hayashi

Osaka University
Nagoya University
Kanazawa University
University of Tokyo

Research output: Contribution to journal › Article › Scientific › peer-review

30 Citations (Scopus)

Abstract

Cellobiose dehydrogenase (CDH) is a dual domain flavocytochrome, which consists of a dehydrogenase (DH) domain containing a flavin adenine dinucleotide and a cytochrome (CYT) domain containing b-type heme. To directly visualize the dynamic domain motion of class-I CDH from Phanerochaete chrysosporium (PcCDH) during catalysis using high-speed atomic force microscopy, the apo-form of PcCDH was anchored to a heme-immobilized flat gold surface that can specifically fix the orientation of the CYT domain. The two domains of CDH are found to be immobile in the absence of cellobiose, whereas the addition of cellobiose triggers an interdomain flip-flop motion involving domain-domain association and dissociation. Our results indicate that dynamic motion of a dual domain enzyme during catalysis induces efficient electron transfer to an external electron acceptor.

Original language	English
Pages (from-to)	6561-6565
Journal	Chemical Science
Volume	8
Issue number	9
DOIs	https://doi.org/10.1039/c7sc01672g
Publication status	Published - 1 Sept 2017
MoE publication type	A1 Journal article-refereed

Funding

This work was supported by JSPS KAKENHI Grant Number JP25102527 and JP15H00746 in Innovative Areas “New Polymeric Materials Based on Element Block” to A.O., JSPS KAKENHI Grant Number JP15H05804 in Innovative Areas “Precisely Designed Catalysts with Customized Scaffolding” to T.H., JSPS KAKENHI Grant Number JP16H00758 in Innovative Areas “Dynamical Ordering & Integrated Functions” and JSPS KAKENHI Grant Number JP16H00830 in Innovative Areas “Soft Molecular Systems” to T.U., and JSPS KAKENHI Grant Number JP24114001 and JP24114008 in Innovative Areas “The Plant Cell Wall as Information Processing System” to K.I. H.H. acknowledges support from the Program for Leading Graduate Schools for Osaka University: Interdisciplinary Program for Biomedical Sciences (IPBS). K.I. acknowledges support from the Asahi Glass Foundation.

Access to Document

10.1039/c7sc01672gLicence: CC BY

Cite this

@article{974b5e557e2d40c3ae5d9bdace36d441,

title = "Interdomain flip-flop motion visualized in flavocytochrome cellobiose dehydrogenase using high-speed atomic force microscopy during catalysis",

abstract = "Cellobiose dehydrogenase (CDH) is a dual domain flavocytochrome, which consists of a dehydrogenase (DH) domain containing a flavin adenine dinucleotide and a cytochrome (CYT) domain containing b-type heme. To directly visualize the dynamic domain motion of class-I CDH from Phanerochaete chrysosporium (PcCDH) during catalysis using high-speed atomic force microscopy, the apo-form of PcCDH was anchored to a heme-immobilized flat gold surface that can specifically fix the orientation of the CYT domain. The two domains of CDH are found to be immobile in the absence of cellobiose, whereas the addition of cellobiose triggers an interdomain flip-flop motion involving domain-domain association and dissociation. Our results indicate that dynamic motion of a dual domain enzyme during catalysis induces efficient electron transfer to an external electron acceptor.",

author = "Hirofumi Harada and Akira Onoda and Takayuki Uchihashi and Hiroki Watanabe and Naoki Sunagawa and Masahiro Samejima and Kiyohiko Igarashi and Takashi Hayashi",

year = "2017",

month = sep,

day = "1",

doi = "10.1039/c7sc01672g",

language = "English",

volume = "8",

pages = "6561--6565",

journal = "Chemical Science",

issn = "2041-6520",

publisher = "Royal Society of Chemistry RSC",

number = "9",

}

TY - JOUR

T1 - Interdomain flip-flop motion visualized in flavocytochrome cellobiose dehydrogenase using high-speed atomic force microscopy during catalysis

AU - Harada, Hirofumi

AU - Onoda, Akira

AU - Uchihashi, Takayuki

AU - Watanabe, Hiroki

AU - Sunagawa, Naoki

AU - Samejima, Masahiro

AU - Igarashi, Kiyohiko

AU - Hayashi, Takashi

PY - 2017/9/1

Y1 - 2017/9/1

N2 - Cellobiose dehydrogenase (CDH) is a dual domain flavocytochrome, which consists of a dehydrogenase (DH) domain containing a flavin adenine dinucleotide and a cytochrome (CYT) domain containing b-type heme. To directly visualize the dynamic domain motion of class-I CDH from Phanerochaete chrysosporium (PcCDH) during catalysis using high-speed atomic force microscopy, the apo-form of PcCDH was anchored to a heme-immobilized flat gold surface that can specifically fix the orientation of the CYT domain. The two domains of CDH are found to be immobile in the absence of cellobiose, whereas the addition of cellobiose triggers an interdomain flip-flop motion involving domain-domain association and dissociation. Our results indicate that dynamic motion of a dual domain enzyme during catalysis induces efficient electron transfer to an external electron acceptor.

AB - Cellobiose dehydrogenase (CDH) is a dual domain flavocytochrome, which consists of a dehydrogenase (DH) domain containing a flavin adenine dinucleotide and a cytochrome (CYT) domain containing b-type heme. To directly visualize the dynamic domain motion of class-I CDH from Phanerochaete chrysosporium (PcCDH) during catalysis using high-speed atomic force microscopy, the apo-form of PcCDH was anchored to a heme-immobilized flat gold surface that can specifically fix the orientation of the CYT domain. The two domains of CDH are found to be immobile in the absence of cellobiose, whereas the addition of cellobiose triggers an interdomain flip-flop motion involving domain-domain association and dissociation. Our results indicate that dynamic motion of a dual domain enzyme during catalysis induces efficient electron transfer to an external electron acceptor.

UR - https://www.scopus.com/pages/publications/85027993264

U2 - 10.1039/c7sc01672g

DO - 10.1039/c7sc01672g

M3 - Article

SN - 2041-6520

VL - 8

SP - 6561

EP - 6565

JO - Chemical Science

JF - Chemical Science

IS - 9

ER -

Interdomain flip-flop motion visualized in flavocytochrome cellobiose dehydrogenase using high-speed atomic force microscopy during catalysis

Abstract

Funding

Access to Document

Other files and links

Fingerprint

Cite this