title: Calcium is essential for the major pseudopilin in the type 2 secretion system.
abstract: The pseudopilus is a key feature of the type 2 secretion system (T2SS) and is made up of multiple pseudopilins that are similar in fold to the type 4 pilins. However, pilins have disulfide bridges, whereas the major pseudopilins of T2SS do not. A key question is therefore how the pseudopilins, and in particular, the most abundant major pseudopilin, GspG, obtain sufficient stability to perform their function. Crystal structures of Vibrio cholerae, Vibrio vulnificus, and enterohemorrhagic Escherichia coli (EHEC) GspG were elucidated, and all show a calcium ion bound at the same site. Conservation of the calcium ligands fully supports the suggestion that calcium ion binding by the major pseudopilin is essential for the T2SS. Functional studies of GspG with mutated calcium ion-coordinating ligands were performed to investigate this hypothesis and show that in vivo protease secretion by the T2SS is severely impaired. Taking all evidence together, this allows the conclusion that, in complete contrast to the situation in the type 4 pili system homologs, in the T2SS, the major protein component of the central pseudopilus is dependent on calcium ions for activity.
authors.affiliation: Department of Biochemistry, Biomolecular Structure Center, University of Washington, Seattle, Washington 98195, USA.
authors.forename: Konstantin V
authors.initials: KV
authors.lastname: Korotkov
authors.forename: Miranda D
authors.initials: MD
authors.lastname: Gray
authors.forename: Allison
authors.initials: A
authors.lastname: Kreger
authors.forename: Stewart
authors.initials: S
authors.lastname: Turley
authors.forename: Maria
authors.initials: M
authors.lastname: Sandkvist
authors.forename: Wim G J
authors.initials: WG
authors.lastname: Hol
mesh_terms: D001692:Biological Transport / Q000502:physiology; D002118:Calcium / Q000737:chemistry* / Q000378:metabolism; D002413:Cations, Divalent / Q000737:chemistry / Q000378:metabolism; D054324:Enterohemorrhagic Escherichia coli / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D036781:Fimbriae Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D008024:Ligands; D009154:Mutation; D011485:Protein Binding / Q000502:physiology; D055550:Protein Stability; D017434:Protein Structure, Tertiary / Q000502:physiology; D040681:Structural Homology, Protein; D013329:Structure-Activity Relationship; D014734:Vibrio cholerae / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism
chemical_list: D002413:Cations, Divalent; D008024:Ligands; D036781:Fimbriae Proteins; D002118:Calcium