ClyJ Is a Novel Pneumococcal Chimeric Lysin with a Cysteine- and Histidine-Dependent Amidohydrolase/Peptidase Catalytic Domain

Yang, Hang; Gong, Yujing; Zhang, Huaidong; Etobayeva, Irina V.; Miernikiewicz, Paulina; Luo, Dehua; Li, Xiaohong; Zhang, Xiaoxu; Dąbrowska, Krystyna; Nelson, Daniel; He, Jin; Wei, Hongping

doi:10.1128/aac.02043-18

Cited by 23 publications

(18 citation statements)

References 58 publications

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“…For example, ClyH was reported to protect 80% of lethal S. aureus challenged mice under a higher dose of 1 mg/mouse [ 33 ]. Using a small dose of lysin for treatment would not only ensure safety because of its dose-dependent cytotoxicity in human cell lines [ 14 ], but also benefit the reduction of immune responses which might lead to side effects. Despite the in vitro increase in bactericidal activity of ClyC in the presence of Ca 2+ , there was no significant difference between the survival rate of the mice treated with ClyC and those treated with ClyC plus 100 μM Ca 2+ .…”

Section: Discussionmentioning

confidence: 99%

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A Highly Active Chimeric Lysin with a Calcium-Enhanced Bactericidal Activity against Staphylococcus aureus In Vitro and In Vivo

Wang

Nyaruaba

et al. 2021

Antibiotics

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Lysins, including chimeric lysins, have recently been explored as novel promising alternatives to failing antibiotics in treating multi-drug resistant (MDR) pathogens, including methicillin-resistant Staphylococcus aureus (MRSA). Herein, by fusing the CHAP (cysteine, histidine-dependent amidohydrolase/peptidase) catalytic domain from the Ply187 lysin with the non-SH3b cell-wall binding domain from the LysSA97 lysin, a new chimeric lysin ClyC was constructed with Ca2+-enhanced bactericidal activity against all S. aureus strains tested, including MRSA. Notably, treating S. aureus with 50 μg/mL of ClyC in the presence of 100 μM Ca2+ lead to a reduction of 9 Log10 (CFU/mL) in viable bacterial number, which was the first time to observe a lysin showing such a high activity. In addition, the effective concentration of ClyC could be decreased dramatically from 12 to 1 μg/mL by combination with 0.3 μg/mL of penicillin G. In a mouse model of S. aureus bacteremia, a single intraperitoneal administration of 0.1 mg/mouse of ClyC significantly improved the survival rates and reduced 2 Log10 (CFU/mL) of the bacterial burdens in the organs of the infected mice. ClyC was also found stable after lyophilization without cryoprotectants. Based on the above observations, ClyC could be a promising candidate against S. aureus infections.

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Section: Discussionmentioning

confidence: 99%

“…Some phage lysins have been reported to have good activities against S. aureus and are in clinical trials now [ 11 , 12 , 13 ]. However, some lysins may have dose-dependent toxicity [ 14 ]. It is important for a lysin drug candidate to possess as high as possible activity in order to have a better therapeutic index.…”

Section: Introductionmentioning

confidence: 99%

A Highly Active Chimeric Lysin with a Calcium-Enhanced Bactericidal Activity against Staphylococcus aureus In Vitro and In Vivo

Wang

Nyaruaba

et al. 2021

Antibiotics

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“…The modular structure of lysins makes it possible to design bioengineered endolysins that have desired properties, such as higher activity, or broader killing spectrum. Chimeric endolysins such as ClyH and ClyJ are engineered to fight against methicillin-resistant Staphylococcus aureus (MRSA) and multidrug-resistant Streptococcus pneumoniae (MRSP) [23,24].…”

Section: Introductionmentioning

confidence: 99%

Structural and functional insights into a novel two-component endolysin encoded by a single gene in Enterococcus faecalis phage

et al. 2020

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Using bacteriophage-derived endolysins as an alternative strategy for fighting drug-resistant bacteria has recently been garnering renewed interest. However, their application is still hindered by their narrow spectra of activity. In our previous work, we demonstrated that the endolysin LysIME-EF1 possesses efficient bactericidal activity against multiple strains of Enterococcus faecalis (E. faecalis). Herein, we observed an 8 kDa fragment and hypothesized that it contributes to LysIME-EF1 lytic activity. To examine our hypothesis, we determined the structure of LysIME-EF1 at 1.75 Å resolution. LysIME-EF1 exhibits a unique architecture in which one full-length LysIME-EF1 forms a tetramer with three additional Cterminal cell-wall binding domains (CBDs) that correspond to the abovementioned 8 kDa fragment. Furthermore, we identified an internal ribosomal binding site (RBS) and alternative start codon within LysIME-EF1 gene, which are demonstrated to be responsible for the translation of the truncated CBD. To elucidate the molecular mechanism for the lytic activity of LysIME-EF1, we combined mutagenesis, lytic activity assays and in vivo animal infection experiments. The results confirmed that the additional LysIME-EF1 CBDs are important for LysIME-EF1 architecture and its lytic activity. To our knowledge, this is the first determined structure of multimeric endolysin encoded by a single gene in E. faecalis phages. As such, it may provide valuable insights into designing potent endolysins against the opportunistic pathogen E. faecalis.

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“…A positively charged CD-only lysin was shown to maintain its lytic activity while possessing a broader lytic spectrum (Low et al, 2011). Bactericidal activity can be improved by fusing the CBD of a streptococcal lysin with the CD of a different lysin, or by modifying the linker between CD and CBD (Yang et al, 2019(Yang et al, , 2020.…”

Section: Introductionmentioning

confidence: 99%

Engineered Lysins With Customized Lytic Activities Against Enterococci and Staphylococci

et al. 2020

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The emergence of multidrug-resistant bacteria has made minor bacterial infections incurable with many existing antibiotics. Lysins are phage-encoded peptidoglycan hydrolases that have demonstrated therapeutic potential as a novel class of antimicrobials. The modular architecture of lysins enables the functional domains – catalytic domain (CD) and cell wall binding domain (CBD) – to be shuffled to create novel lysins. The CD is classically thought to be only involved in peptidoglycan hydrolysis whereas the CBD dictates the lytic spectrum of a lysin. While there are many studies that extended the lytic spectrum of a lysin by domain swapping, few have managed to introduce species specificity in a chimeric lysin. In this work, we constructed two chimeric lysins by swapping the CBDs of two parent lysins with different lytic spectra against enterococci and staphylococci. We showed that these chimeric lysins exhibited customized lytic spectra distinct from the parent lysins. Notably, the chimeric lysin P10N-V12C, which comprises a narrow-spectrum CD fused with a broad-spectrum CBD, displayed species specificity not lysing Enterococcus faecium while targeting Enterococcus faecalis and staphylococci. Such species specificity can be attributed to the narrow-spectrum CD of the chimeric lysin. Using flow cytometry and confocal microscopy, we found that the E. faecium cells that were treated with P10N-V12C are less viable with compromised membranes yet remained morphologically intact. Our results suggest that while the CBD is a major determinant of the lytic spectrum of a lysin, the CD is also responsible in the composition of the final lytic spectrum, especially when it pertains to species-specificity.

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ClyJ Is a Novel Pneumococcal Chimeric Lysin with a Cysteine- and Histidine-Dependent Amidohydrolase/Peptidase Catalytic Domain

Cited by 23 publications

References 58 publications

A Highly Active Chimeric Lysin with a Calcium-Enhanced Bactericidal Activity against Staphylococcus aureus In Vitro and In Vivo

A Highly Active Chimeric Lysin with a Calcium-Enhanced Bactericidal Activity against Staphylococcus aureus In Vitro and In Vivo

Structural and functional insights into a novel two-component endolysin encoded by a single gene in Enterococcus faecalis phage

Engineered Lysins With Customized Lytic Activities Against Enterococci and Staphylococci

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