The Transition from Closed to Open Conformation of Treponema pallidum Outer Membrane-associated Lipoprotein TP0453 Involves Membrane Sensing and Integration by Two Amphipathic Helices

Luthra, Amit; Zhu, Guangyu; Desrosiers, Daniel C.; Eggers, Christian H.; Mulay, Vishwaroop; Anand, Asha; McArthur, Fiona; Romano, Fabian B.; Caimano, Melissa J.; Heuck, Alejandro P.; Malkowski, Michael G.; Radolf, Justin D.

doi:10.1074/jbc.m111.305284

Cited by 24 publications

(33 citation statements)

References 56 publications

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“…Pore formation assays were performed using large unilamellar vesicles (LUVs) loaded with the fluorophore Tb(DPA) 3 3Ϫ as previously described (40,48). Immunologic reagents.…”

Section: Methodsmentioning

confidence: 99%

The Major Outer Sheath Protein (Msp) of Treponema denticola Has a Bipartite Domain Architecture and Exists as Periplasmic and Outer Membrane-Spanning Conformers

et al. 2013

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“…Pore formation assays were performed using large unilamellar vesicles (LUVs) loaded with the fluorophore Tb(DPA) 3 3Ϫ as previously described (40,48). Immunologic reagents.…”

Section: Methodsmentioning

confidence: 99%

The Major Outer Sheath Protein (Msp) of Treponema denticola Has a Bipartite Domain Architecture and Exists as Periplasmic and Outer Membrane-Spanning Conformers

et al. 2013

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“…After protein purification, the protein was concentrated using Amicon-Ultra centrifugal filters (EMD Millipore, Billerica, MA). Treponema pallidum protein TP0453 and E. coli OmpF were purified as described elsewhere (49,50).…”

Section: Methodsmentioning

confidence: 99%

“…Pore formation assays were performed as described previously (50,62). Liposomes loaded with Tb(DPA) 3 3Ϫ were diluted in 50 mM Tris (pH 7.5), 100 mM NaCl supplemented with 5 mM EDTA to a concentration of 100 M total lipids.…”

Section: ϫmentioning

confidence: 99%

Structural Modeling and Physicochemical Characterization Provide Evidence that P66 Forms a β-Barrel in the Borrelia burgdorferi Outer Membrane

et al. 2014

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The Borrelia burgdorferi outer membrane (OM) contains numerous surface-exposed lipoproteins but a relatively low density of integral OM proteins (OMPs). Few membrane-spanning OMPs of B. burgdorferi have been definitively identified, and none are well characterized structurally. Here, we provide evidence that the borrelial OMP P66, a known adhesin with pore-forming activity, forms a ␤-barrel in the B. burgdorferi OM. Multiple computer-based algorithms predict that P66 forms a ␤-barrel with either 22 or 24 transmembrane domains. According to our predicted P66 topology, a lysine residue (K487) known to be sensitive to trypsin cleavage is located within a surface-exposed loop. When we aligned the mature P66 amino acid sequences from B. burgdorferi and B. garinii, we found that K487 was present only in the B. burgdorferi P66 protein sequence. When intact cells from each strain were treated with trypsin, only B. burgdorferi P66 was trypsin sensitive, indicating that K487 is surface exposed, as predicted. Consistent with this observation, when we inserted a c-Myc tag adjacent to K487 and utilized surface localization immunofluorescence, we detected the loop containing K487 on the surface of B. burgdorferi. P66 was examined by both Triton X-114 phase partitioning and circular dichroism, confirming that the protein is amphiphilic and contains extensive (48%) ␤-sheets, respectively. Moreover, P66 also was able to incorporate into liposomes and form channels in large unilamellar vesicles. Finally, blue native PAGE (BN-PAGE) revealed that under nondenaturing conditions, P66 is found in large complexes of ϳ400 kDa and ϳ600 kDa. Outer surface lipoprotein A (OspA) and OspB both coimmunoprecipitate with P66, demonstrating that P66 associates with OspA and OspB in B. burgdorferi. The combined computer-based structural analyses and supporting physicochemical properties of P66 provide a working model to further examine the porin and integrin-binding activities of this OMP as they relate to B. burgdorferi physiology and Lyme disease pathogenesis.

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“…The mechanism(s) by which the parasite acquires and utilizes these essential nutrients can potentially help explain the peculiar membrane biology of T. pallidum, elucidate key aspects of its parasitic strategy, and prompt new avenues of investigation for potentially novel antimicrobial drug targets. To this end, we have found that many of the likely transport systems of T. pallidum are predicated on the organism's membrane lipoproteins (5)(6)(7)(8)(9)(10)(11)(12)(13), and, as a result, the organism devotes a large percentage of its genome to encoding these lipoproteins (3,14,15). Unfortunately, many of the putative lipoproteins are hypothetical (i.e.…”

mentioning

confidence: 99%

The TP0796 Lipoprotein of Treponema pallidum Is a Bimetal-dependent FAD Pyrophosphatase with a Potential Role in Flavin Homeostasis

Deka

Brautigam

Liu

et al. 2013

Journal of Biological Chemistry

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Background:The TP0796 lipoprotein of Treponema pallidum belongs to the poorly characterized ApbE superfamily. Results: TP0796 hydrolyzed FAD into FMN and AMP, consistent with the general enzymatic mechanism of an FAD pyrophosphatase. Conclusion: This novel metal-dependent enzyme probably plays an essential role in flavin homeostasis in T. pallidum. Significance: This is the first description of a metal-dependent FAD pyrophosphatase in bacteria.

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The Transition from Closed to Open Conformation of Treponema pallidum Outer Membrane-associated Lipoprotein TP0453 Involves Membrane Sensing and Integration by Two Amphipathic Helices

Cited by 24 publications

References 56 publications

The Major Outer Sheath Protein (Msp) of Treponema denticola Has a Bipartite Domain Architecture and Exists as Periplasmic and Outer Membrane-Spanning Conformers

The Major Outer Sheath Protein (Msp) of Treponema denticola Has a Bipartite Domain Architecture and Exists as Periplasmic and Outer Membrane-Spanning Conformers

Structural Modeling and Physicochemical Characterization Provide Evidence that P66 Forms a β-Barrel in the Borrelia burgdorferi Outer Membrane

The TP0796 Lipoprotein of Treponema pallidum Is a Bimetal-dependent FAD Pyrophosphatase with a Potential Role in Flavin Homeostasis

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