title: AglJ adds the first sugar of the N-linked pentasaccharide decorating the Haloferax volcanii S-layer glycoprotein.
abstract: Like the Eukarya and Bacteria, the Archaea also perform N glycosylation. Using the haloarchaeon Haloferax volcanii as a model system, a series of Agl proteins involved in the archaeal version of this posttranslational modification has been identified. In the present study, the participation of HVO_1517 in N glycosylation was considered, given its homology to a known component of the eukaryal N-glycosylation pathway and because of the genomic proximity of HVO_1517 to agl genes encoding known elements of the H. volcanii N-glycosylation process. By combining the deletion of HVO_1517 with mass spectrometric analysis of both dolichol phosphate monosaccharide-charged carriers and the S-layer glycoprotein, evidence was obtained showing the participation of HVO_1517, renamed AglJ, in adding the first hexose of the N-linked pentasaccharide decorating this reporter glycoprotein. The deletion of aglJ, however, did not fully prevent the attachment of a hexose residue to the S-layer glycoprotein. Moreover, in the absence of AglJ, the level of only one of the three monosaccharide-charged dolichol phosphate carriers detected in the cell was reduced. Nonetheless, in cells lacking AglJ, no further sugar subunits were added to the remaining monosaccharide-charged dolichol phosphate carriers or to the monosaccharide-modified S-layer glycoprotein, pointing to the importance of the sugar added through the actions of AglJ for proper N glycosylation. Finally, while aglJ can be deleted, H. volcanii surface layer integrity is compromised in the absence of the encoded protein.
authors.affiliation: Dept. of Life Sciences, Ben Gurion University, Beersheva 84105, Israel.
authors.forename: Lina
authors.initials: L
authors.lastname: Kaminski
authors.forename: Mehtap
authors.initials: M
authors.lastname: Abu-Qarn
authors.forename: Ziqiang
authors.initials: Z
authors.lastname: Guan
authors.forename: Shai
authors.initials: S
authors.lastname: Naparstek
authors.forename: Valeria V
authors.initials: VV
authors.lastname: Ventura
authors.forename: Christian R H
authors.initials: CR
authors.lastname: Raetz
authors.forename: Paul G
authors.initials: PG
authors.lastname: Hitchen
authors.forename: Anne
authors.initials: A
authors.lastname: Dell
authors.forename: Jerry
authors.initials: J
authors.lastname: Eichler
mesh_terms: D019843:Archaeal Proteins / Q000235:genetics / Q000378:metabolism*; D050260:Carbohydrate Metabolism; D002352:Carrier Proteins / Q000378:metabolism; D017353:Gene Deletion; D019848:Gene Expression Regulation, Archaeal / Q000502:physiology*; D006031:Glycosylation; D019622:Haloferax volcanii / Q000235:genetics / Q000378:metabolism*; D006601:Hexoses / Q000378:metabolism; D008562:Membrane Glycoproteins / Q000378:metabolism*; D008969:Molecular Sequence Data; D015394:Molecular Structure
chemical_list: D019843:Archaeal Proteins; D002352:Carrier Proteins; D006601:Hexoses; D008562:Membrane Glycoproteins; C055764:S-layer proteins
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