title: Regulation of a phage endolysin by disulfide caging.
abstract: In contrast to canonical phage endolysins, which require holin-mediated disruption of the membrane to gain access to attack the cell wall, signal anchor release (SAR) endolysins are secreted by the host sec system, where they accumulate in an inactive form tethered to the membrane by their N-terminal SAR domains. SAR endolysins become activated by various mechanisms upon release from the membrane. In its inactive form, the prototype SAR endolysin, Lyz(P1), of coliphage P1, has an active-site Cys covalently blocked by a disulfide bond; activation involves a disulfide bond isomerization driven by a thiol in the newly released SAR domain, unblocking the active-site Cys. Here, we report that Lyz(103), the endolysin of Erwinia phage ERA103, is also a SAR endolysin. Although Lyz(103) does not have a catalytic Cys, genetic evidence suggests that it also is activated by a thiol-disulfide isomerization triggered by a thiol in the SAR domain. In this case, the inhibitory disulfide in nascent Lyz(103) is formed between cysteine residues flanking a catalytic glutamate, caging the active site. Thus, Lyz(P1) and Lyz(103) define subclasses of SAR endolysins that differ in the nature of their inhibitory disulfide, and Lyz(103) is the first enzyme found to be regulated by disulfide bond caging of its active site.
authors.affiliation: 2128 TAMU, Department of Biochemistry and Biophysics, Texas A&M University, College Station, TX 77843-2128, USA.
authors.forename: Gabriel F
authors.initials: GF
authors.lastname: Kuty
authors.forename: Min
authors.initials: M
authors.lastname: Xu
authors.forename: Douglas K
authors.initials: DK
authors.lastname: Struck
authors.forename: Elizabeth J
authors.initials: EJ
authors.lastname: Summer
authors.forename: Ry
authors.initials: R
authors.lastname: Young
mesh_terms: D000595:Amino Acid Sequence; D001435:Bacteriophages / Q000378:metabolism*; D004220:Disulfides / Q000737:chemistry*; D010450:Endopeptidases / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D044084:Erwinia amylovora / Q000821:virology*; D004926:Escherichia coli; D015967:Gene Expression Regulation, Viral / Q000502:physiology*; D008969:Molecular Sequence Data; D017434:Protein Structure, Tertiary; D014764:Viral Proteins / Q000235:genetics / Q000378:metabolism
chemical_list: D004220:Disulfides; D014764:Viral Proteins; D010450:Endopeptidases; C001143:endolysin