Characterization of multiprotein complexes of the Borrelia burgdorferi outer membrane vesicles.

Xiuli Yang, Kamoltip Promnares, Jinhong Qin, Ming He, Deborah Y Shroder, Toru Kariu, Yan Wang, Utpal Pal

Department of Veterinary Medicine, University of Maryland , College Park and Virginia-Maryland Regional College of Veterinary Medicine, United States.

Journal of proteome research 2011 Oct 7

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Among bacterial cell envelopes, the Borrelia burgdorferi outer membrane (OM) is structurally unique in that the identities of many protein complexes remain unknown; however, their characterization is the first step toward our understanding of membrane protein interactions and potential functions. Here, we used two-dimensional blue native/SDS-PAGE/mass spectrometric analysis for a global characterization of protein-protein interactions as well as to identify protein complexes in OM vesicles isolated from multiple infectious sensu stricto isolates of B. burgdorferi. Although we uncovered the existence of at least 10 distinct OM complexes harboring several unique subunits, the complexome is dominated by the frequent occurrence of a limited diversity of membrane proteins, most notably P13, outer surface protein (Osp) A, -B, -C, and -D and Lp6.6. The occurrence of these complexes and specificity of subunit interaction were further supported by independent two-dimensional immunoblotting and coimmunoprecipitation assays as well as by mutagenesis studies, where targeted depletion of a subunit member (P66) selectively abolished a specific complex. Although a comparable profile of the OM complexome was detected in two major infectious isolates, such as B31 and 297, certain complexes are likely to occur in an isolate-specific manner. Further assessment of protein complexes in multiple Osp-deficient isolates showed loss of several protein complexes but revealed the existence of additional complex/subunits that are undetectable in wild-type cells. Together, these observations uncovered borrelial antigens involved in membrane protein interactions. The study also suggests that the assembly process of OM complexes is specific and that the core or stabilizing subunits vary between complexes. Further characterization of these protein complexes including elucidation of their biological significance may shed new light on the mechanism of pathogen persistence and the development of preventative measures against the infection.

Citation

Xiuli Yang, Kamoltip Promnares, Jinhong Qin, Ming He, Deborah Y Shroder, Toru Kariu, Yan Wang, Utpal Pal. Characterization of multiprotein complexes of the Borrelia burgdorferi outer membrane vesicles. Journal of proteome research. 2011 Oct 7;10(10):4556-66