Structural insights into the mechanism of Escherichia coli YmdB: A 2'-O-acetyl-ADP-ribose deacetylase.
The Escherichia coli protein YmdB belongs to the macrodomain protein family, which can bind ADP-ribose (ADPr) and its derivatives. Recently, YmdB was reported to be capable of deacetylating O-acetyl-ADP-ribose (OAADPr) to yield ADPr and free acetate. To study the substrate specificity and catalytic mechanism, the crystal structures of E. coli YmdB in complex with ADPr, double mutant N25AD35A complexed with 2'-OAADPr, and Y126A/ADPr complex were solved at 1.8Å, 2.8Å and 3.0Å resolution, respectively. Structural and biochemical studies reveal that YmdB has substrate specificity against 2'-OAADPr. The conserved residues Asn25 and Asp35 are crucial for catalytic activity, and an active water molecule is proposed as the nucleophile to attack the acetyl group of 2'-OAADPr. Our findings indicate that the conserved phenyl group of Tyr126 plays a crucial role in catalytic activity by stabilizing the right orientation of distal ribose and that Gly32 may be important for activity by interacting with the acetyl group of 2'-OAADPr. Based on these observations, a model of YmdB in complex with 2'-OAADPr was made to illustrate the proposed catalytic mechanism of YmdB.
Hefei National Laboratory for Physical Sciences at Microscale, School of Life Sciences, University of Science and Technology of China, 96 Jinzhai Road, Hefei, Anhui 230026, People's Republic of China; Key Laboratory of Structural Biology, Chinese Academy of Sciences, Hefei, Anhui 230027, People's Republic of China; National Synchrotron Radiation Laboratory, University of Science and Technology of China, Hefei, Anhui 230027, People's Republic of China.
Weichang
W
Zhang
Hefei National Laboratory for Physical Sciences at Microscale, School of Life Sciences, University of Science and Technology of China, 96 Jinzhai Road, Hefei, Anhui 230026, People's Republic of China; Key Laboratory of Structural Biology, Chinese Academy of Sciences, Hefei, Anhui 230027, People's Republic of China; National Synchrotron Radiation Laboratory, University of Science and Technology of China, Hefei, Anhui 230027, People's Republic of China.
Chengliang
C
Wang
Hefei National Laboratory for Physical Sciences at Microscale, School of Life Sciences, University of Science and Technology of China, 96 Jinzhai Road, Hefei, Anhui 230026, People's Republic of China; Key Laboratory of Structural Biology, Chinese Academy of Sciences, Hefei, Anhui 230027, People's Republic of China; National Synchrotron Radiation Laboratory, University of Science and Technology of China, Hefei, Anhui 230027, People's Republic of China.
Yang
Y
Song
Hefei National Laboratory for Physical Sciences at Microscale, School of Life Sciences, University of Science and Technology of China, 96 Jinzhai Road, Hefei, Anhui 230026, People's Republic of China; Key Laboratory of Structural Biology, Chinese Academy of Sciences, Hefei, Anhui 230027, People's Republic of China; National Synchrotron Radiation Laboratory, University of Science and Technology of China, Hefei, Anhui 230027, People's Republic of China.
Chen
C
Shao
Hefei National Laboratory for Physical Sciences at Microscale, School of Life Sciences, University of Science and Technology of China, 96 Jinzhai Road, Hefei, Anhui 230026, People's Republic of China; Key Laboratory of Structural Biology, Chinese Academy of Sciences, Hefei, Anhui 230027, People's Republic of China; National Synchrotron Radiation Laboratory, University of Science and Technology of China, Hefei, Anhui 230027, People's Republic of China. Electronic address: xuanzbin@ustc.edu.cn.
Xuan
X
Zhang
Hefei National Laboratory for Physical Sciences at Microscale, School of Life Sciences, University of Science and Technology of China, 96 Jinzhai Road, Hefei, Anhui 230026, People's Republic of China; Key Laboratory of Structural Biology, Chinese Academy of Sciences, Hefei, Anhui 230027, People's Republic of China; National Synchrotron Radiation Laboratory, University of Science and Technology of China, Hefei, Anhui 230027, People's Republic of China. Electronic address: zangjy@ustc.edu.cn.
Jianye
J
Zang
D000246:Adenosine Diphosphate Ribose / Q000737:chemistry; D000595:Amino Acid Sequence; D002352:Carrier Proteins / Q000235:genetics / Q000502:physiology* / Q000648:ultrastructure*; D002384:Catalysis; D018360:Crystallography, X-Ray; D004926:Escherichia coli / Q000201:enzymology* / Q000235:genetics; D029968:Escherichia coli Proteins / Q000235:genetics / Q000502:physiology* / Q000648:ultrastructure*; D006867:Hydrolases; D008958:Models, Molecular; D008969:Molecular Sequence Data; D009154:Mutation / Q000235:genetics; D038961:O-Acetyl-ADP-Ribose / Q000737:chemistry*; D011485:Protein Binding / Q000502:physiology; D017434:Protein Structure, Tertiary; D013379:Substrate Specificity / Q000502:physiology
C449679:2-O-acetyl-ADP-ribose; D002352:Carrier Proteins; D029968:Escherichia coli Proteins; D038961:O-Acetyl-ADP-Ribose; C534806:YmdB protein, E coli; D000246:Adenosine Diphosphate Ribose; D006867:Hydrolases
ADPr; Catalytic mechanism; Deacetylase; Macrodomain; OAADPr; YmdB