A Novel Marker for the Species-Specific Detection and Quantitation of Vibrio cholerae by Targeting an Outer Membrane Lipoprotein lolB Gene

2013 ◽  
Vol 23 (4) ◽  
pp. 555-559 ◽  
Author(s):  
Min Seok Cho
Keyword(s):  
Outer Membrane ◽  
Vibrio Cholerae ◽  
Specific Detection ◽  
Outer Membrane Lipoprotein ◽  
Membrane Lipoprotein ◽  
Species Specific

scholarly journals Identification of a TcpC-TcpQ Outer Membrane Complex Involved in the Biogenesis of the Toxin-Coregulated Pilus of Vibrio cholerae

2005 ◽  
Vol 187 (7) ◽  
pp. 2225-2232 ◽  
Author(s):  
Niranjan Bose ◽  
Ronald K. Taylor
Keyword(s):  
Outer Membrane ◽  
Vibrio Cholerae ◽  
Sodium Dodecyl ◽  
Direct Interaction ◽  
High Molecular Weight Complex ◽  
Metal Affinity ◽  
Membrane Complex ◽  
Outer Membrane Lipoprotein ◽  
Membrane Lipoprotein ◽  
Toxin Coregulated Pilus

ABSTRACT The toxin-coregulated pilus (TCP) of Vibrio cholerae and the soluble TcpF protein that is secreted via the TCP biogenesis apparatus are essential for intestinal colonization. The TCP biogenesis apparatus is composed of at least nine proteins but is largely uncharacterized. TcpC is an outer membrane lipoprotein required for TCP biogenesis that is a member of the secretin protein superfamily. In the present study, analysis of TcpC in a series of strains deficient in each of the TCP biogenesis proteins revealed that TcpC was absent specifically in a tcpQ mutant. TcpQ is a predicted periplasmic protein required for TCP biogenesis. Fractionation studies revealed that the protein is not localized to the periplasm but is associated predominantly with the outer membrane fraction. An analysis of the amount of TcpQ present in the series of tcp mutants demonstrated the inverse of the TcpC result (absence of TcpQ in a tcpC deletion strain). Complementation of the tcpQ deletion restored TcpC levels and TCP formation, and similarly, complementation of tcpC restored TcpQ. Metal affinity pull-down experiments performed using His-tagged TcpC or TcpQ demonstrated a direct interaction between TcpC and TcpQ. In the presence of TcpQ, TcpC was found to form a high-molecular-weight complex that is stable in 2% sodium dodecyl sulfate and at temperatures below 65°C, a characteristic of secretin complexes. Fractionation studies in which TcpC was overexpressed in the absence of TcpQ showed that TcpQ is also required for proper localization of TcpC to the outer membrane.

Backbone resonance assignments of the outer membrane lipoprotein FrpD from Neisseria meningitidis

2012 ◽  
Vol 8 (1) ◽  
pp. 53-55 ◽  
Author(s):  
Ladislav Bumba ◽  
Ekaterina Sviridova ◽  
Ivana Kutá Smatanová ◽  
Pavlína Řezáčová ◽  
Václav Veverka
Keyword(s):  
Neisseria Meningitidis ◽  
Outer Membrane ◽  
Resonance Assignments ◽  
Backbone Resonance Assignments ◽  
Backbone Resonance ◽  
Outer Membrane Lipoprotein ◽  
Membrane Lipoprotein

scholarly journals Role of the Pilot Protein YscW in the Biogenesis of the YscC Secretin in Yersinia enterocolitica

2004 ◽  
Vol 186 (16) ◽  
pp. 5366-5375 ◽  
Author(s):  
Peter Burghout ◽  
Frank Beckers ◽  
Emmie de Wit ◽  
Ria van Boxtel ◽  
Guy R. Cornelis ◽  
...  
Keyword(s):  
Outer Membrane ◽  
Yersinia Enterocolitica ◽  
Amino Acid Residues ◽  
Wild Type ◽  
C Terminus ◽  
Outer Membrane Lipoprotein ◽  
Protein Secretion System ◽  
Membrane Lipoprotein ◽  
Type Iii Protein Secretion

ABSTRACT The YscC secretin is a major component of the type III protein secretion system of Yersinia enterocolitica and forms an oligomeric structure in the outer membrane. In a mutant lacking the outer membrane lipoprotein YscW, secretion is strongly reduced, and it has been proposed that YscW plays a role in the biogenesis of the secretin. To study the interaction between the secretin and this putative pilot protein, YscC and YscW were produced in trans in a Y. enterocolitica strain lacking all other components of the secretion machinery. YscW expression increased the yield of oligomeric YscC and was required for its outer membrane localization, confirming the function of YscW as a pilot protein. Whereas the pilot-binding site of other members of the secretin family has been identified in the C terminus, a truncated YscC derivative lacking the C-terminal 96 amino acid residues was functional and stabilized by YscW. Pulse-chase experiments revealed that ∼30 min were required before YscC oligomerization was completed. In the absence of YscW, oligomerization was delayed and the yield of YscC oligomers was strongly reduced. An unlipidated form of the YscW protein was not functional, although it still interacted with the secretin and caused mislocalization of YscC even in the presence of wild-type YscW. Hence, YscW interacts with the unassembled YscC protein and facilitates efficient oligomerization, likely at the outer membrane.

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