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BioMetals

, Volume 32, Issue 2, pp 317–328 | Cite as

Molecular structure of thermostable and zinc-ion-binding γ-class carbonic anhydrases

  • Wenming WangEmail author
  • Yao Zhang
  • Lele Wang
  • Qi Jing
  • Xiaolu Wang
  • Xiaoli Xi
  • Xuan Zhao
  • Hongfei WangEmail author
Article
  • 97 Downloads

Abstract

The γ-class carbonic anhydrases (γ-CAs) mainly come from methanogens methane-producing bacteria that grow in hot springs and catalyze the interconversion of carbon dioxide and water to bicarbonate and protons. Here, the γ-CA from Thermus thermophilus HB8 (γ-TtCA) was expressed and purified, its crystal structure was determined at 2.3 Å resolution in space group P1. The asymmetric unit contains two trimers and six catalytic Zn2+. In general, the fold of the protein is similar to those of homologous enzymes from Geobacillus Kaustophilus, Bacillus Cereus, Methanosarcina Thermophila and others. Each monomer comprises a triangular prism-like structure consisting of a left-handed β-helix and a C-terminal α-helix. The catalytic Zn2+ bound to three histidines and a phosphate radical in a tetrahedral fashion. It is located at the interface between the two monomers. Inductively coupled plasma mass spectrometry measurements further suggest that the molar ratio of zinc ions and protein molecules is 1:1. The structure revealed a novel different region situated between the left-handed β-helix and the C-terminal α-helix. Compared to previously reported structures, half of the C-terminal α-helix was replaced with a long loop in this structure. The purified γ-TtCA exhibits no significant carbonic anhydrase activity compared to α-class carbonic anhydrases. This study provides insight into the structural diversity of γ-CAs with potential function for γ-CAs.

Keywords

γ-Class carbonic anhydrases Thermus thermophilus HB8 Crystal structure 

Notes

Acknowledgements

The work was supported by the National Natural Science Foundation of China (Nos. 21601112 and 21671125), Talent Plan of Shanxi Province and Shanxi Scholarship Council of China (2015-021). We thank the staff at the BL17U1/18U1/19U1 beamline of National Center for Protein Sciences Shanghai (NCPSS) at Shanghai Synchrotron Radiation Facility for assistance during data collection. Crystallographic coordinates of γ-TtCAΔ169 are deposited to the Protein Data Bank with PDB code 6IVE.

Compliance with ethical standards

Conflict of interest

All of the authors of our manuscript declare that we have no competing interests.

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Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.Key Laboratory of Chemical Biology and Molecular Engineering of Education Ministry, Institute of Molecular ScienceShanxi UniversityTaiyuanChina
  2. 2.Department of ChemistryUniversity of MemphisMemphisUSA

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