title: Archaeal JAB1/MPN/MOV34 metalloenzyme (HvJAMM1) cleaves ubiquitin-like small archaeal modifier proteins (SAMPs) from protein-conjugates.
abstract: Proteins with JAB1/MPN/MOV34 metalloenzyme (JAMM/MPN+) domains are widespread among all domains of life, yet poorly understood. Here we report the purification and characterization of an archaeal JAMM/MPN+ domain protein (HvJAMM1) from Haloferax volcanii that cleaves ubiquitin-like small archaeal modifier proteins (SAMP1/2) from protein conjugates. HvJAMM1 cleaved SAMP1/2 conjugates generated in H. volcanii as well as isopeptide- and linear-linked SAMP1-MoaE in purified form. Cleavage of linear linked SAMP1-MoaE was dependent on the presence of the SAMP domain and the C-terminal VSGG motif of this domain. While HvJAMM1 was inhibited by size exclusion chromatography and metal chelators, its activity could be restored by addition of excess ZnCl2 . HvJAMM1 residues (Glu31, His88, His90, Ser98 and Asp101) that were conserved with the JAMM/MPN+ active-site motif were required for enzyme activity. Together, these results provide the first example of a JAMM/MPN+ zinc metalloprotease that independently catalyses the cleavage of ubiquitin-like (isopeptide and linear) bonds from target proteins. In archaea, HvJAMM1 likely regulates sampylation and the pools of 'free' SAMP available for protein modification. HvJAMM1-type proteins are thought to release the SAMPs from proteins modified post-translationally as well as those synthesized as domain fusions.
authors.affiliation: Department of Microbiology and Cell Science, University of Florida, Gainesville, FL 32611-0700, USA.
authors.forename: Nathaniel L
authors.initials: NL
authors.lastname: Hepowit
authors.forename: Sivakumar
authors.initials: S
authors.lastname: Uthandi
authors.forename: Hugo V
authors.initials: HV
authors.lastname: Miranda
authors.forename: Micaela
authors.initials: M
authors.lastname: Toniutti
authors.forename: Laurence
authors.initials: L
authors.lastname: Prunetti
authors.forename: Oliver
authors.initials: O
authors.lastname: Olivarez
authors.forename: Ian Mitchelle S
authors.initials: IM
authors.lastname: De Vera
authors.forename: Gail E
authors.initials: GE
authors.lastname: Fanucci
authors.forename: Sixue
authors.initials: S
authors.lastname: Chen
authors.forename: Julie A
authors.initials: JA
authors.lastname: Maupin-Furlow
mesh_terms: D019843:Archaeal Proteins / Q000302:isolation & purification / Q000378:metabolism*; D002712:Chlorides / Q000378:metabolism; D004252:DNA Mutational Analysis; D020536:Enzyme Activators / Q000378:metabolism; D019622:Haloferax volcanii / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D008666:Metalloendopeptidases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D050505:Mutant Proteins / Q000235:genetics / Q000378:metabolism; D020125:Mutation, Missense; D059748:Proteolysis; D025841:Small Ubiquitin-Related Modifier Proteins / Q000378:metabolism*; D017967:Zinc Compounds / Q000378:metabolism
chemical_list: D019843:Archaeal Proteins; D002712:Chlorides; D020536:Enzyme Activators; D050505:Mutant Proteins; D025841:Small Ubiquitin-Related Modifier Proteins; D017967:Zinc Compounds; C016837:zinc chloride; D008666:Metalloendopeptidases