Bacteriophage protein PEIP is a potent Bacillus subtilis enolase inhibitor

Kaining Zhang; Shanshan Li; Yawen Wang; Zhihao Wang; Nancy Mulvenna; Hang Yang; Peipei Zhang; Huan Chen; Yan Li; Hongliang Wang; Yongxiang Gao; Sivaramesh Wigneshweraraj; Steve Matthews; Kaiming Zhang; Bing Liu

doi:10.1016/j.celrep.2022.111026

Bacteriophage protein PEIP is a potent Bacillus subtilis enolase inhibitor

Cell Rep. 2022 Jul 5;40(1):111026. doi: 10.1016/j.celrep.2022.111026.

Authors

Kaining Zhang¹, Shanshan Li², Yawen Wang³, Zhihao Wang⁴, Nancy Mulvenna⁵, Hang Yang⁶, Peipei Zhang¹, Huan Chen¹, Yan Li⁷, Hongliang Wang⁶, Yongxiang Gao², Sivaramesh Wigneshweraraj⁵, Steve Matthews⁸, Kaiming Zhang⁹, Bing Liu¹⁰

Affiliations

¹ BioBank, The First Affiliated Hospital of Xi'an Jiaotong University, Shaanxi 710061, China; Institute for Protein Science and Phage Research, The First Affiliated Hospital of Xi'an Jiaotong University, Shaanxi 710061, China.
² MOE Key Laboratory for Cellular Dynamics and School of Life Sciences, University of Science and Technology of China, Hefei 230027, China.
³ BioBank, The First Affiliated Hospital of Xi'an Jiaotong University, Shaanxi 710061, China; Institute for Protein Science and Phage Research, The First Affiliated Hospital of Xi'an Jiaotong University, Shaanxi 710061, China; Department of Laboratory Medicine, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi, 710061, China.
⁴ BioBank, The First Affiliated Hospital of Xi'an Jiaotong University, Shaanxi 710061, China; Institute for Protein Science and Phage Research, The First Affiliated Hospital of Xi'an Jiaotong University, Shaanxi 710061, China; MRC Centre for Molecular Bacteriology and Infection, Imperial College London, London, UK.
⁵ MRC Centre for Molecular Bacteriology and Infection, Imperial College London, London, UK.
⁶ Department of Pathogen Biology and Immunology, Xi'an Jiaotong University Health Science Center, Xi'an, Shaanxi 710061, China.
⁷ Department of Pathogen Biology, School of Basic Medicine, Tongji Medical College, Huazhong University of Science and Technology, 13 Hangkong Road, Wuhan, Hubei 430030, China.
⁸ Institute for Protein Science and Phage Research, The First Affiliated Hospital of Xi'an Jiaotong University, Shaanxi 710061, China; MRC Centre for Molecular Bacteriology and Infection, Imperial College London, London, UK.
⁹ MOE Key Laboratory for Cellular Dynamics and School of Life Sciences, University of Science and Technology of China, Hefei 230027, China. Electronic address: kmzhang@ustc.edu.cn.
¹⁰ BioBank, The First Affiliated Hospital of Xi'an Jiaotong University, Shaanxi 710061, China; Institute for Protein Science and Phage Research, The First Affiliated Hospital of Xi'an Jiaotong University, Shaanxi 710061, China; Department of Laboratory Medicine, The First Affiliated Hospital of Xi'an Jiaotong University, Xi'an, Shaanxi, 710061, China. Electronic address: bliu2018@xjtu.edu.cn.

PMID: 35793626
DOI: 10.1016/j.celrep.2022.111026

Abstract

Enolase is a highly conserved enzyme that presents in all organisms capable of glycolysis or fermentation. Its immediate product phosphoenolpyruvate is essential for other important processes like peptidoglycan synthesis and the phosphotransferase system in bacteria. Therefore, enolase inhibitors are of great interest. Here, we report that Gp60, a phage-encoded enolase inhibitor protein (PEIP) of bacteriophage SPO1 for Bacillus subtilis, is an enolase inhibitor. PEIP-expressing bacteria exhibit growth attenuation, thinner cell walls, and safranin color in Gram staining owing to impaired peptidoglycan synthesis. We solve the structure of PEIP-enolase tetramer and show that PEIP disassembles enolase by disrupting the basic dimer unit. The structure reveals that PEIP does not compete for substrate binding but induces a cascade of conformational changes that limit accessibility to the enolase catalytic site. This phage-inspired disassembly of enolase represents an alternative strategy for the development of anti-microbial drugs.

Keywords: CP: Microbiology; bacteriophage; enolase inhibitor; glycolysis; peptidoglycan; phosphoenolpyruvate; single-particle cryo-EM.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Bacillus subtilis* / metabolism
Bacteriophages* / metabolism
Catalytic Domain
Peptidoglycan / metabolism
Phosphopyruvate Hydratase / metabolism

Substances

Peptidoglycan
Phosphopyruvate Hydratase