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Video Abstract (AI generated) (01:53) Paper PreprintMulti-drug resistance (MDR) is a serious threat to global public health, making the development of new antimicrobials an urgent necessity. Pyocins are protein antibiotics produced by Pseudomonas aeruginosa strains to kill closely related cells during intraspecific competition. Here, we report an in depth biochemical, microbicidal and structural characterization of a new S-type pyocin, named S8. Initially, we described the domain organization and secondary structure of S8. Subsequently, we observed that a recombinant S8 composed of the killing subunit in complex with the immunity (Im) protein killed the strain PAO1. Furthermore, mutation of a highly conserved glutamic acid to alanine (Glu100Ala) completely inhibited this antimicrobial activity. Probably the integrity of the H-N-H motif is essential in the killing activity of S8, as Glu100 is a highly conserved component of this structure. Next, we observed that S8 is a metal-dependent endonuclease, as EDTA treatment abolished its ability to cleave supercoiled pUC18 plasmid. Supplementation of apo S8 with Ni2+ strongly induced this DNase activity, whereas Mn2+ and Mg2+ exhibited moderate effects and Zn2+ was inhibitory. Additionally, S8 bound Zn2+ with a higher affinity than Ni2+ and the Glu100Ala mutation decreased the affinity of S8 for these metals as shown by isothermal titration calorimetry (ITC). Finally, we describe the crystal structure of the Glu100Ala pyocin-S8DNase-Im complex at 1.38 , which gave us new insights into the endonuclease activity of S8. Our results reinforce the possible use of S8 as an alternative antibiotic for MDR Pseudomonas aeruginosa strains, while leaving commensal human microbiota intact.
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Nilton Lincopan. (2021, Nov 3).Structural and functional analysis of pyocin S8 from Pseudomonas aeruginosa : requirement of a glutamate in the H-N-H motif for the DNase activity[Video]. Scitok. https://scitok.com/project/p/17580b06
G. Turano Helena. "Structural and functional analysis of pyocin S8 from Pseudomonas aeruginosa : requirement of a glutamate in the H-N-H motif for the DNase activity" Scitok, uploaded by Lincopan Nilton, 3 Nov, 2021, https://scitok.com/project/p17580b06
Nilton Lincopan. "Structural and functional analysis of pyocin S8 from Pseudomonas aeruginosa : requirement of a glutamate in the H-N-H motif for the DNase activity" Scitok. (Nov 3, 2021). https://scitok.com/project/p/17580b06
Nilton Lincopan (Nov 3, 2021). Structural and functional analysis of pyocin S8 from Pseudomonas aeruginosa : requirement of a glutamate in the H-N-H motif for the DNase activity Scitok. https://scitok.com/project/p/17580b06
Nilton Lincopan. Structural and functional analysis of pyocin S8 from Pseudomonas aeruginosa : requirement of a glutamate in the H-N-H motif for the DNase activity[video]. 2021 Nov 3. https://scitok.com/project/p/17580b06
@online{al2006link, title={ Structural and functional analysis of pyocin S8 from Pseudomonas aeruginosa : requirement of a glutamate in the H-N-H motif for the DNase activity }, author={ Lincopan, Nilton }, organization={Scitok}, month={ Nov }, day={ 3 }, year={ 2021 }, url = {https://scitok.com/project/p/17580b06}, }