Structural and functional insights into Skl and Pal endolysins, two cysteine-amidases with anti-pneumococcal activity. Dithiothreitol (DTT) effect on lytic activity
| Autor: | Palma Rico-Lastres, Rubén M. Buey, J. Fernando Díaz, Margarita Menéndez, Guadalupe García, Pedro García, Noelia Hernández-Ortiz, Cristina Gallego-Páramo |
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| Přispěvatelé: | Ministerio de Economía y Competitividad (España), Ministerio de Ciencia e Innovación (España), Agencia Estatal de Investigación (España), Centro de Investigación Biomédica en Red Enfermedades Respiratorias (España), Instituto de Salud Carlos III |
| Rok vydání: | 2021 |
| Předmět: |
Microbiology (medical)
Lysis Chemistry Endolysin In silico CHAP domain Lysin Anti-pneumococcal activity Cysteine-peptidase Microbiology DTT-mediated activation QR1-502 Dithiothreitol Homology (biology) Amidase chemistry.chemical_compound Biochemistry Reducing agents Choline-binding domain Amidase_5 domain Original Research Cysteine |
| Zdroj: | Digital.CSIC. Repositorio Institucional del CSIC instname 'Frontiers in Microbiology ', vol: 12, pages: 740914-1-740914-20 (2021) Frontiers in Microbiology, Vol 12 (2021) Frontiers in Microbiology |
| ISSN: | 1664-302X |
| Popis: | 20 pags, 9 figs, 2 tabs. -- The Supplementary Material for this article can be found online at: https://www.frontiersin.org/articles/10.3389/fmicb.2021.740914/full#supplementary-material We have structurally and functionally characterized Skl and Pal endolysins, the latter being the first endolysin shown to kill effectively Streptococcus pneumoniae, a leading cause of deathly diseases. We have proved that Skl and Pal are cysteine-amidases whose catalytic domains, from CHAP and Amidase_5 families, respectively, share an α3β6-fold with papain-like topology. Catalytic triads are identified (for the first time in Amidase_5 family), and residues relevant for substrate binding and catalysis inferred from in silico models, including a calcium-binding site accounting for Skl dependence on this cation for activity. Both endolysins contain a choline-binding domain (CBD) with a β-solenoid fold (homology modeled) and six conserved choline-binding loci whose saturation induced dimerization. Remarkably, Pal and Skl dimers display a common overall architecture, preserved in choline-bound dimers of pneumococcal lysins with other catalytic domains and bond specificities, as disclosed using small angle X-ray scattering (SAXS). Additionally, Skl is proved to be an efficient anti-pneumococcal agent that kills multi-resistant strains and clinical emergent-serotype isolates. Interestingly, Skl and Pal time-courses of pneumococcal lysis were sigmoidal, which might denote a limited access of both endolysins to target bonds at first stages of lysis. Furthermore, their DTT-mediated activation, of relevance for other cysteine-peptidases, cannot be solely ascribed to reversal of catalytic-cysteine oxidation. This work was supported by grants from the Ministry of Economy and Competitiveness (BFU2015-70072-R) and the Ministry of Science, Innovation and Universities (RTI2018-099985-B-I00/AEI/10.13039/501100011033) to MM, and by a grant from the Ministry of Economy and Competitiveness (MINECO-FEDER, SAF2017-88664-R) to PG. Additional funding was provided by the Centro de Investigacion Biomedica en Red de Enfermedades Respiratorias (CIBERES), an initiative of the Instituto de Salud Carlos III (ISCIII), to MM and PG. |
| Databáze: | OpenAIRE |
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