Role of Net Charge on Catalytic Domain and Influence of Cell Wall Binding Domain on Bactericidal Activity, Specificity, and Host Range of Phage Lysins

Low, Lieh Yoon; Yang, Chen; Perego, Marta; Osterman, Andrei L.; Liddington, Robert

doi:10.1074/jbc.m111.244160

Cited by 131 publications

(144 citation statements)

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“…Alternatively, it could be that the CD27L cell wall binding activity is not fully effective when added externally; since it has evolved to act from inside the cell, it may not be able to access the peptidoglycan or its binding target from the outside due to external structures, such as the extensive array of surface layer proteins. It has been proposed that endolysin C-terminal domains exhibit an inhibitory activity when not bound to the correct target (27,28). Although the presence of the CD27L C-terminal domain did not reduce activity of the CS74L catalytic domain on C. sporogenes, it did have a negative effect on CSCD lytic activity against C. tyrobutyricum.…”

Section: Discussionmentioning

confidence: 81%

Genomic Sequence of Bacteriophage ATCC 8074-B1 and Activity of Its Endolysin and Engineered Variants against Clostridium sporogenes

Mayer

Gasson

Narbad

2012

Appl Environ Microbiol

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ABSTRACTLytic bacteriophage ATCC 8074-B1 produces large plaques on its hostClostridium sporogenes. Sequencing of the 47,595-bp genome allowed the identification of 82 putative open reading frames, including those encoding proteins for head and tail morphogenesis and lysis. However, sequences commonly associated with lysogeny were absent. ORF 22 encodes an endolysin, CS74L, that shows homology toN-acetylmuramoyl-l-alanine amidases, and when expressed inEscherichia coli, the protein causes effective lysis ofC. sporogenescells when added externally. CS74L was also active onClostridium tyrobutyricumandClostridium acetobutylicum. The catalytic domain expressed alone (CS74L1–177) exhibited a similar activity and the same host range as the full-length endolysin. A chimeric endolysin consisting of the CS74L catalytic domain fused to the C-terminal domain of endolysin CD27L, derived fromClostridium difficilebacteriophage ΦCD27, was produced. This chimera (CSCD) lysedC. sporogenescells with an activity equivalent to that of the catalytic domain alone. In contrast, the CD27L C-terminal domain reduced the efficacy of the CS74L catalytic domain when tested againstC. tyrobutyricum. The addition of the CD27L C-terminal domain did not enable the lysin to targetC. difficileor other CD27L-sensitive bacteria.

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Section: Discussionmentioning

confidence: 81%

Genomic Sequence of Bacteriophage ATCC 8074-B1 and Activity of Its Endolysin and Engineered Variants against Clostridium sporogenes

Mayer

Gasson

Narbad

2012

Appl Environ Microbiol

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“…It has been proposed that the cell wall binding domain of endolysin has an inhibitory role when it is not bound to the target (58,62). However, this is not the case for LysPBC1, because the cell wall binding domain of LysPBC1 did not bind several B. cereus group strains (our unpublished data) despite the fact that they were more sensitive to the full-length endolysin.…”

Section: Resultsmentioning

confidence: 43%

Bacteriophage PBC1 and Its Endolysin as an Antimicrobial Agent against Bacillus cereus

Kong

Ryu

2015

Appl Environ Microbiol

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Bacillus cereus is an opportunistic human pathogen responsible for food poisoning and other, nongastrointestinal infections. Due to the emergence of multidrug-resistant B. cereus strains, the demand for alternative therapeutic options is increasing. To address these problems, we isolated and characterized a Siphoviridae virulent phage, PBC1, and its lytic enzymes. PBC1 showed a very narrow host range, infecting only 1 of 22 B. cereus strains. Phylogenetic analysis based on the major capsid protein revealed that PBC1 is more closely related to the Bacillus clarkii phage BCJA1c and phages of lactic acid bacteria than to the phages infecting B. cereus. Whole-genome comparison showed that the late-gene region, including the terminase gene, structural genes, and holin gene of PBC1, is similar to that from B. cereus temperate phage 250, whereas their endolysins are different. Compared to the extreme host specificity of PBC1, its endolysin, LysPBC1, showed a much broader lytic spectrum, albeit limited to the genus Bacillus. The catalytic domain of LysPBC1 when expressed alone also showed Bacillus-specific lytic activity, which was lower against the B. cereus group but higher against the Bacillus subtilis group than the full-length protein. Taken together, these results suggest that the virulent phage PBC1 is a useful component of a phage cocktail to control B. cereus, even with its exceptionally narrow host range, as it can kill a strain of B. cereus that is not killed by other phages, and that LysPBC1 is an alternative biocontrol agent against B. cereus. Bacillus cereus is a spore-forming, opportunistic Gram-positive bacterium that is widely distributed in the environment. Because B. cereus produces emetic toxin and enterotoxins, it causes food poisoning, including vomiting and diarrhea, particularly after the consumption of rice-based dishes (1). The Centers for Disease and Control and Prevention (CDC) reported that B. cereus outbreaks account for 2 to 5% of food-borne diseases in the United States (2). In addition to food poisoning, B. cereus is also associated with potentially fatal non-gastrointestinal tract infections due to the various types of toxins, including phospholipases, proteases, and hemolysins, produced during growth (3). For these reasons, there is an urgent need to control B. cereus. Since B. cereus is generally resistant to beta-lactam antibiotics and several strains of B. cereus are also resistant to erythromycin, tetracycline, and fluoroquinolones (1, 4), the demand for alternative methods to control B. cereus has grown (3).Bacteriophages, natural killers of bacteria, have gained increasing attention in the past few decades, particularly in light of emerging antibiotic resistance (5, 6). Bacteriophages are much more specific than antibiotics, so they have minimal impact on commensal bacteria other than the host. Although bacteria can develop resistance to their viral predators, finding a new phage that can kill resistant bacteria is not difficult, because the phage itself continually evolves again...

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“…Intramolecular interaction as the cause of inactivation of endolysin in Bacillus anthracis phage was suggested earlier also (42). However, after performing differential scanning calorimetry experiment that showed no interaction between the catalytic domain and the cell wall binding domain, the hypothesis was discarded (49). Here, we demonstrate by SPR that such an interaction does exist between NTD and CTD of D29 Lysin A. Post-translational modifications have also been suggested to activate lysins in other bacteriophages such as E. coli T4 tail lysozyme, P1 Lyz, and streptococcal bacteriophage C 1 PlyC (50 -52).…”

Section: Discussionmentioning

confidence: 99%

Molecular Dissection of Phage Endolysin

Pohane

Joshi

Jain

2014

Journal of Biological Chemistry

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Background: Bacteriophages produce endolysins to hydrolyze host cell wall for their progeny release. Results: Mycobacterium phage endolysin is inactive in E. coli but is active in mycobacteria. Conclusion: An interdomain interaction makes Lysin A inactive in E. coli. Significance: A hydrolase with built-in mechanism attains host specificity. An in-depth study can help in developing strong anti-tuberculosis agents.

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Role of Net Charge on Catalytic Domain and Influence of Cell Wall Binding Domain on Bactericidal Activity, Specificity, and Host Range of Phage Lysins

Cited by 131 publications

References 93 publications

Genomic Sequence of Bacteriophage ATCC 8074-B1 and Activity of Its Endolysin and Engineered Variants against Clostridium sporogenes

Genomic Sequence of Bacteriophage ATCC 8074-B1 and Activity of Its Endolysin and Engineered Variants against Clostridium sporogenes

Bacteriophage PBC1 and Its Endolysin as an Antimicrobial Agent against Bacillus cereus

Molecular Dissection of Phage Endolysin

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