title: Specificity of archaeal caspase activity in the extreme halophile Haloferax volcanii.
abstract: Caspase-like proteases are key initiators and executioners of programmed cell death (PCD), which is initiated by environmental stimuli and manifests in organisms ranging from unicellular microbes to higher eukaryotes. Archaea had been absent from the caspase inheritance discussion due to a lack of gene homologues. We recently demonstrated extremely high, basal caspase-like catalytic activity in the model haloarcheon, Haloferax volcanii, which was linked to the cellular stress response and was widespread among diverse Archaea. Here, we rigorously tested the catalytic specificity of the observed archaeal caspase-like activities using hydrolytic assays with a diverse suite of protease substrates and inhibitors compared with known model serine and cysteine proteases (trypsin, cathepsin, papain, and human caspase-8). Our experiments demonstrate that exponentially growing H. volcanii possesses a highly specific caspase-like activity that most closely resembles caspase-4, is preferentially inhibited by the pancaspase inhibitor, zVAD-FMK, and has no crossreactivity with other known protease families. Our findings firmly root the extremely high levels of caspase-like activity as the dominant proteolytic activity in this extreme haloarcheaon, thereby providing further support for housekeeping functions in Haloarchaea. Given the deep archaeal roots of eukaryotes, we suggest that this activity served as a foundation for stress pathways in higher organisms.
authors.affiliation: Institute for Marine and Coastal Sciences, Rutgers University, New Brunswick, NJ, USA.
authors.forename: Mansha
authors.initials: M
authors.lastname: Seth-Pasricha
authors.forename: Kelly A
authors.initials: KA
authors.lastname: Bidle
authors.forename: Kay D
authors.initials: KD
authors.lastname: Bidle
mesh_terms: D019843:Archaeal Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D020169:Caspases / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism*; D004795:Enzyme Stability; D019622:Haloferax volcanii / Q000737:chemistry / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D006801:Humans; D012965:Sodium Chloride / Q000378:metabolism; D013045:Species Specificity; D013379:Substrate Specificity
chemical_list: D019843:Archaeal Proteins; D012965:Sodium Chloride; D020169:Caspases
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