Structural and biochemical characterization reveals LysGH15 as an unprecedented 'EF-hand-like' calcium-binding phage lysin
| العنوان: | Structural and biochemical characterization reveals LysGH15 as an unprecedented 'EF-hand-like' calcium-binding phage lysin |
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| المؤلفون: | Changjiang Sun, Chongtao Du, Ziyin Cui, Fengfeng Niu, Xin Feng, Yue Li, Liancheng Lei, Jingmin Gu, Yingang Feng, Wei Ding, Lianying Jiao, Jun Song, Mei Yang, Songying Ouyang, Dong Han, Yongjun Yang, Wenyu Han, Zhi-Jie Liu, Xiaohe Liu |
| المصدر: | PLoS Pathogens, Vol 10, Iss 5, p e1004109 (2014) PLoS Pathogens |
| بيانات النشر: | Public Library of Science (PLoS), 2014. |
| سنة النشر: | 2014 |
| مصطلحات موضوعية: | Methicillin-Resistant Staphylococcus aureus, Models, Molecular, lcsh:Immunologic diseases. Allergy, Staphylococcus, Immunology, CHAP domain, Protein domain, Molecular Sequence Data, Lysin, Microbial Sensitivity Tests, Microbiology, Amidohydrolases, Protein structure, Mucoproteins, Cell Wall, Virology, Microbial Control, Catalytic Domain, Genetics, Protein Interaction Domains and Motifs, Amino Acid Sequence, Molecular Biology, Microbial Pathogens, lcsh:QH301-705.5, biology, Sequence Homology, Amino Acid, EF hand, Gram Positive Bacteria, Active site, Biology and Life Sciences, Bacteriology, Protein superfamily, Bacterial Pathogens, Anti-Bacterial Agents, Lytic cycle, lcsh:Biology (General), Medical Microbiology, biology.protein, Biophysics, Parasitology, Calcium, Staphylococcus Phages, lcsh:RC581-607, Research Article |
| الوصف: | The lysin LysGH15, which is derived from the staphylococcal phage GH15, demonstrates a wide lytic spectrum and strong lytic activity against methicillin-resistant Staphylococcus aureus (MRSA). Here, we find that the lytic activity of the full-length LysGH15 and its CHAP domain is dependent on calcium ions. To elucidate the molecular mechanism, the structures of three individual domains of LysGH15 were determined. Unexpectedly, the crystal structure of the LysGH15 CHAP domain reveals an “EF-hand-like” calcium-binding site near the Cys-His-Glu-Asn quartet active site groove. To date, the calcium-binding site in the LysGH15 CHAP domain is unique among homologous proteins, and it represents the first reported calcium-binding site in the CHAP family. More importantly, the calcium ion plays an important role as a switch that modulates the CHAP domain between the active and inactive states. Structure-guided mutagenesis of the amidase-2 domain reveals that both the zinc ion and E282 are required in catalysis and enable us to propose a catalytic mechanism. Nuclear magnetic resonance (NMR) spectroscopy and titration-guided mutagenesis identify residues (e.g., N404, Y406, G407, and T408) in the SH3b domain that are involved in the interactions with the substrate. To the best of our knowledge, our results constitute the first structural information on the biochemical features of a staphylococcal phage lysin and represent a pivotal step forward in understanding this type of lysin. Author Summary The staphylococcal phage lysin LysGH15 demonstrates great potential against methicillin-resistant Staphylococcus aureus (MRSA). Here, we report that the lytic activity of LysGH15 and its CHAP domain is dependent on calcium ions. To elucidate the molecular mechanism, we determined the structures of three individual LysGH15 domains using X-ray crystallography or nuclear magnetic resonance (NMR). The crystal structure unexpectedly reveals an “EF-hand-like” calcium-binding site near the Cys-His-Glu-Asn quartet active site groove in the LysGH15 CHAP domain. Furthermore, the calcium ion plays an important role as a switch that modulates the lytic activity of the CHAP domain. Additionally, structure-guided mutagenesis also confirms that both E282 and the zinc ion play an important role in maintaining the lytic activity of the LysGH15 amidase-2 domain. Moreover, the NMR structure and titration-guided mutagenesis identify residues in the LysGH15 SH3b domain that are involved in the interactions with the substrate. The structure of LysGH15 is the first determined lysin structure from a staphylococcal phage, and these results represent a pivotal step forward in understanding this type of lysin. |
| اللغة: | English |
| تدمد: | 1553-7374 1553-7366 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::6c883824609beb87375e2e7e2c1bdd87 http://europepmc.org/articles/PMC4022735?pdf=render |
| Rights: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....6c883824609beb87375e2e7e2c1bdd87 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 15537374 15537366 |
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