title: A Hybrid Mechanism for the Synechocystis Arsenate Reductase Revealed by Structural Snapshots during Arsenate Reduction.
abstract: Evolution of enzymes plays a crucial role in obtaining new biological functions for all life forms. Arsenate reductases (ArsC) are several families of arsenic detoxification enzymes that reduce arsenate to arsenite, which can subsequently be extruded from cells by specific transporters. Among these, the Synechocystis ArsC (SynArsC) is structurally homologous to the well characterized thioredoxin (Trx)-coupled ArsC family but requires the glutaredoxin (Grx) system for its reactivation, therefore classified as a unique Trx/Grx-hybrid family. The detailed catalytic mechanism of SynArsC is unclear and how the "hybrid" mechanism evolved remains enigmatic. Herein, we report the molecular mechanism of SynArsC by biochemical and structural studies. Our work demonstrates that arsenate reduction is carried out via an intramolecular thiol-disulfide cascade similar to the Trx-coupled family, whereas the enzyme reactivation step is diverted to the coupling of the glutathione-Grx pathway due to the local structural difference. The current results support the hypothesis that SynArsC is likely a molecular fossil representing an intermediate stage during the evolution of the Trx-coupled ArsC family from the low molecular weight protein phosphotyrosine phosphatase (LMW-PTPase) family.
authors.affiliation: From the College of Chemistry and Molecular Engineering, Beijing Nuclear Magnetic Resonance Center.
authors.forename: Cuiyun
authors.initials: C
authors.lastname: Hu
authors.affiliation: From the College of Chemistry and Molecular Engineering, Beijing Nuclear Magnetic Resonance Center.
authors.forename: Caifang
authors.initials: C
authors.lastname: Yu
authors.affiliation: From the College of Chemistry and Molecular Engineering.
authors.forename: Yanhua
authors.initials: Y
authors.lastname: Liu
authors.affiliation: From the College of Chemistry and Molecular Engineering, Beijing Nuclear Magnetic Resonance Center.
authors.forename: Xianhui
authors.initials: X
authors.lastname: Hou
authors.affiliation: From the College of Chemistry and Molecular Engineering.
authors.forename: Xiaoyun
authors.initials: X
authors.lastname: Liu
authors.affiliation: From the College of Chemistry and Molecular Engineering, Beijing Nuclear Magnetic Resonance Center, yunfei@pku.edu.cn.
authors.forename: Yunfei
authors.initials: Y
authors.lastname: Hu
authors.affiliation: From the College of Chemistry and Molecular Engineering, Beijing Nuclear Magnetic Resonance Center, College of Life Sciences, Beijing National Laboratory for Molecular Sciences, Peking University, Beijing 100871, China changwen@pku.edu.cn.
authors.forename: Changwen
authors.initials: C
authors.lastname: Jin
mesh_terms: D000595:Amino Acid Sequence; D053502:Arsenate Reductases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D001149:Arsenates / Q000737:chemistry / Q000378:metabolism*; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D001665:Binding Sites; D004220:Disulfides / Q000378:metabolism; D054477:Glutaredoxins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D005978:Glutathione / Q000378:metabolism; D008958:Models, Molecular; D008969:Molecular Sequence Data; D010084:Oxidation-Reduction; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D017386:Sequence Homology, Amino Acid; D013438:Sulfhydryl Compounds / Q000378:metabolism; D046939:Synechocystis / Q000201:enzymology* / Q000235:genetics; D013879:Thioredoxins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
chemical_list: D001149:Arsenates; D001426:Bacterial Proteins; D004220:Disulfides; D054477:Glutaredoxins; D013438:Sulfhydryl Compounds; D013879:Thioredoxins; D053502:Arsenate Reductases; D005978:Glutathione; C025657:arsenic acid
keywords: ArsC; SynArsC; arsenate reductase; arsenic detoxification; enzyme; enzyme catalysis; enzyme mechanism; nuclear magnetic resonance (NMR); oxidation-reduction (redox); protein structure