Previous Article | Next Article 
Infection and Immunity, May 1999, p. 2266-2276, Vol. 67, No. 5
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Membrane Topology and Cellular Location of the Treponema
pallidum Glycerophosphodiester Phosphodiesterase (GlpQ)
Ortholog
Dmitriy V.
Shevchenko,1
Timothy J.
Sellati,1
David L.
Cox,2
Olga V.
Shevchenko,1
Esther J.
Robinson,1 and
Justin
D.
Radolf1,3,*
Departments of Internal
Medicine1 and
Microbiology,3 University of Texas
Southwestern Medical Center, Dallas, Texas 75235, and
Division of STD Laboratory Research, Centers for Disease
Control and Prevention, Atlanta, Georgia 303332
Received 13 November 1998/Returned for modification 7 January
1999/Accepted 1 February 1999
Recent reports that isolated Treponema pallidum outer
membranes contain an ortholog for glycerophosphodiester
phosphodiesterase (GlpQ) (D. V. Shevchenko, D. R. Akins,
E. J. Robinson, M. Li, O. V. Shevchenko, and J. D. Radolf, Infect. Immun. 65:4179-4189, 1997) and that this protein is a
potential opsonic target for T. pallidum (C. E. Stebeck, J. M. Shaffer, T. W. Arroll, S. A. Lukehart,
and W. C. Van Voorhis, FEMS Microbiol. Lett. 154:303-310, 1997)
prompted a more detailed investigation of its physicochemical properties and cellular location. [14C]palmitate
radiolabeling studies of a GlpQ-alkaline phosphatase fusion expressed
in Escherichia coli confirmed the prediction from DNA
sequencing that the protein is lipid modified. Studies using
Triton X-114 phase partitioning revealed that the protein's amphiphilicity is due to lipid modification and that a
substantial portion of the polypeptide is associated with the
T. pallidum peptidoglycan sacculus. Three different
approaches, i.e., (i) proteinase K treatment of intact treponemes,
(ii) indirect immunofluorescence analysis of treponemes encapsulated in
agarose beads, and (iii) opsonophagocytosis of treponemes incubated
with antiserum against recombinant GlpQ by rabbit peritoneal
macrophages, confirmed that GlpQ is entirely subsurface in T. pallidum. Moreover, rabbits hyperimmunized with GlpQ were not
protected against intradermal challenge with virulent
treponemes. Circular dichroism spectroscopy confirmed that the
recombinant form of the polypeptide lacked discernible evidence of
denaturation. Finally, GlpQ was not radiolabeled when T. pallidum outer membranes were incubated with
3-(trifluoromethyl)-3-(m-[125I]iodophenyl)-diazarene,
a photoactivatable, lipophilic probe which promiscuously labels both
proteins and lipids within phospholipid bilayers. Taken as a
whole, these studies indicate that the T. pallidum GlpQ
ortholog is a periplasmic protein associated predominantly with the
spirochete's peptidoglycan-cytoplasmic membrane complex.
*
Corresponding author. Present address: Center for
Microbial Pathogenesis, University of Connecticut Health Center, 263 Farmington Ave., Farmington, CT 06030-3710. Phone: (860) 679-8129. Fax:
(860) 679-8130. E-mail: JRadolf{at}up.uchl.edu.
Infection and Immunity, May 1999, p. 2266-2276, Vol. 67, No. 5
0019-9567/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
This article has been cited by other articles:
-
Toropainen, M., Raitolehto, A., Henckaerts, I., Wauters, D., Poolman, J., Lestrate, P., Kayhty, H.
(2008). Pneumococcal Haemophilus influenzae Protein D Conjugate Vaccine Induces Antibodies That Inhibit Glycerophosphodiester Phosphodiesterase Activity of Protein D. Infect. Immun.
76: 4546-4553
[Abstract]
[Full Text]
-
Brinkman, M. B., McKevitt, M., McLoughlin, M., Perez, C., Howell, J., Weinstock, G. M., Norris, S. J., Palzkill, T.
(2006). Reactivity of Antibodies from Syphilis Patients to a Protein Array Representing the Treponema pallidum Proteome.. J. Clin. Microbiol.
44: 888-891
[Abstract]
[Full Text]
-
LaFond, R. E., Lukehart, S. A.
(2006). Biological Basis for Syphilis. Clin. Microbiol. Rev.
19: 29-49
[Abstract]
[Full Text]
-
Hazlett, K. R. O., Cox, D. L., Decaffmeyer, M., Bennett, M. P., Desrosiers, D. C., La Vake, C. J., La Vake, M. E., Bourell, K. W., Robinson, E. J., Brasseur, R., Radolf, J. D.
(2005). TP0453, a Concealed Outer Membrane Protein of Treponema pallidum, Enhances Membrane Permeability. J. Bacteriol.
187: 6499-6508
[Abstract]
[Full Text]
-
Champion, C. I., Blanco, D. R., Lovett, M. A.
(2005). Quantitative Assessment of Protection in Experimental Syphilis. Infect. Immun.
73: 5923-5927
[Abstract]
[Full Text]
-
Van Voorhis, W. C., Barrett, L. K., Lukehart, S. A., Schmidt, B., Schriefer, M., Cameron, C. E.
(2003). Serodiagnosis of Syphilis: Antibodies to Recombinant Tp0453, Tp92, and Gpd Proteins Are Sensitive and Specific Indicators of Infection by Treponema pallidum. J. Clin. Microbiol.
41: 3668-3674
[Abstract]
[Full Text]
-
McKevitt, M., Patel, K., Smajs, D., Marsh, M., McLoughlin, M., Norris, S. J., Weinstock, G. M., Palzkill, T.
(2003). Systematic Cloning of Treponema pallidum Open Reading Frames for Protein Expression and Antigen Discovery. Genome Res
13: 1665-1674
[Abstract]
[Full Text]
-
Schwan, T. G., Battisti, J. M., Porcella, S. F., Raffel, S. J., Schrumpf, M. E., Fischer, E. R., Carroll, J. A., Stewart, P. E., Rosa, P., Somerville, G. A.
(2003). Glycerol-3-Phosphate Acquisition in Spirochetes: Distribution and Biological Activity of Glycerophosphodiester Phosphodiesterase (GlpQ) among Borrelia Species. J. Bacteriol.
185: 1346-1356
[Abstract]
[Full Text]
-
Matsunaga, J., Young, T. A., Barnett, J. K., Barnett, D., Bolin, C. A., Haake, D. A.
(2002). Novel 45-Kilodalton Leptospiral Protein That Is Processed to a 31-Kilodalton Growth-Phase-Regulated Peripheral Membrane Protein. Infect. Immun.
70: 323-334
[Abstract]
[Full Text]
-
Hazlett, K. R.O., Sellati, T. J., Nguyen, T. T., Cox, D. L., Clawson, M. L., Caimano, M. J., Radolf, J. D.
(2001). The Tprk Protein of Treponema pallidum Is Periplasmic and Is Not a Target of Opsonic Antibody or Protective Immunity. JEM
193: 1015-1026
[Abstract]
[Full Text]
-
Cox, D. L., Radolf, J. D.
(2001). Insertion of fluorescent fatty acid probes into the outer membranes of the pathogenic spirochaetes Treponema pallidum and Borrelia burgdorferi. Microbiology
147: 1161-1169
[Abstract]
[Full Text]
-
Haake, D. A.
(2000). Spirochaetal lipoproteins and pathogenesis. Microbiology
146: 1491-1504
[Full Text]
-
Centurion-Lara, A., Godornes, C., Castro, C., Van Voorhis, W. C., Lukehart, S. A.
(2000). The tprK Gene Is Heterogeneous among Treponema pallidum Strains and Has Multiple Alleles. Infect. Immun.
68: 824-831
[Abstract]
[Full Text]
-
Sellati, T. J., Bouis, D. A., Caimano, M. J., Feulner, J. A., Ayers, C., Lien, E., Radolf, J. D.
(1999). Activation of Human Monocytic Cells by Borrelia burgdorferi and Treponema pallidum Is Facilitated by CD14 and Correlates with Surface Exposure of Spirochetal Lipoproteins. J. Immunol.
163: 2049-2056
[Abstract]
[Full Text]
-
Deka, R. K., Lee, Y.-H., Hagman, K. E., Shevchenko, D., Lingwood, C. A., Hasemann, C. A., Norgard, M. V., Radolf, J. D.
(1999). Physicochemical Evidence that Treponema pallidum TroA Is a Zinc-Containing Metalloprotein That Lacks Porin-Like Structure. J. Bacteriol.
181: 4420-4423
[Abstract]
[Full Text]