Existence of Separate Domains in Lysin PlyG for Recognizing Bacillus anthracis Spores and Vegetative Cells

Abstract

<jats:title>ABSTRACT</jats:title><jats:p>As a potential antimicrobial, the bacteriophage lysin PlyG has been reported to specifically recognize<jats:named-content xmlns:xlink="http://www.w3.org/1999/xlink" content-type="genus-species" xlink:type="simple">Bacillus anthracis</jats:named-content>vegetative cells only and to kill<jats:named-content xmlns:xlink="http://www.w3.org/1999/xlink" content-type="genus-species" xlink:type="simple">B. anthracis</jats:named-content>vegetative cells and its germinating spores. However, how PlyG interacts with<jats:named-content xmlns:xlink="http://www.w3.org/1999/xlink" content-type="genus-species" xlink:type="simple">B. anthracis</jats:named-content>spores remains unclear. Herein, a 60-amino-acid domain in PlyG (residues 106 to 165), located mainly in the previously identified catalytic domain, was found able to specifically recognize<jats:named-content xmlns:xlink="http://www.w3.org/1999/xlink" content-type="genus-species" xlink:type="simple">B. anthracis</jats:named-content>spores but not vegetative cells. The exosporium of the spores was found to be the most probable binding target of this domain. This is the first time that a lysin for spore-forming bacteria has been found to have separate domains to recognize spores and vegetative cells, which might help in understanding the coevolution of phages with spore-forming bacteria. Besides providing new biomarkers for developing better assays for identifying<jats:named-content xmlns:xlink="http://www.w3.org/1999/xlink" content-type="genus-species" xlink:type="simple">B. anthracis</jats:named-content>spores, the newly found domain may be helpful in developing PlyG as a preventive antibiotic to reduce the threat of anthrax in suspected exposures to<jats:named-content xmlns:xlink="http://www.w3.org/1999/xlink" content-type="genus-species" xlink:type="simple">B. anthracis</jats:named-content>spores.</jats:p>

Antimicrobial bacteriophage-derived proteins and therapeutic applications

“…This endolysin has multiple binding domains, one for peptidoglycan of vegetative cells and another for the spore surface. 40 In contrast, CBD are not always required for lytic activity, 31,41 and in some cases CBD removal can improve bacteriolytic potency and/or lytic spectrum (see 'Engineering novel endolysin constructs' section below for more details). Endolysins are typically the product of a single gene in a single open reading frame.…”

Novel Chimeric Lysin with High-Level Antimicrobial Activity against Methicillin-Resistant Staphylococcus aureus In Vitro and In Vivo

“…The recombinant proteins were expressed by the E. coli BL21(DE3) strain in standard LB medium and purified following procedures described previously (24,29), with minor modifications. ClyH was induced overnight in BL21(DE3) cells with L-arabinose in a final concentration of 0.2% at 20°C.…”

Sensitive and rapid detection of staphylococcus aureus in milk via cell binding domain of lysin

“…The CBDs of the majority of lysins are responsible for attaching the hydrolases to their specific substrates in the bacterial cell walls via non-covalent binding of carbohydrate ligands (Loessner et al, 2002). It has been reported that the number of CBD binding sites on one bacterial cell can be 10 7 or more (Yang et al, 2012). Moreover, the size of CBD (usually 10 to 20 kilodaltons) is much smaller than that of antibodies (usually 150 kilodaltons).…”

“…Bacteriophage endolysins or lysins are a class of bacteria cell wall peptidoglycan hydrolase synthesized at the late stage of the bacteriophage lytic cycle, mediating lysis of the host cell and the release of progeny phages (Fischetti, 2008;Yang et al, 2012Yang et al, , 2014b domain and C-terminal cell wall binding domain (CBD) (Yang et al, 2012(Yang et al, , 2014b. The CBDs of the majority of lysins are responsible for attaching the hydrolases to their specific substrates in the bacterial cell walls via non-covalent binding of carbohydrate ligands (Loessner et al, 2002).…”