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Journal of Bacteriology, July 1999, p. 4420-4423, Vol. 181, No. 14
Departments of
Microbiology1 and Internal
Medicine,2 The University of Texas
Southwestern Medical Center, Dallas, Texas 75235, and the
Division of Research Microbiology, Research Institute,
Hospital for Sick Children, Toronto, Ontario M5G 1X8,
Canada3
Received 30 November 1998/Accepted 3 May 1999
Although TroA (Tromp1) was initially reported to be a
Treponema pallidum outer membrane protein with porin-like
properties, subsequent studies have suggested that it actually is a
periplasmic substrate-binding protein involved in the transport of
metals across the treponemal cytoplasmic membrane. Here we conducted additional physicochemical studies to address the divergent viewpoints concerning this protein. Triton X-114 phase partitioning of recombinant TroA constructs with or without a signal sequence corroborated our
prior contention that the native protein's amphiphilic behavior is due
to its uncleaved leader peptide. Whereas typical porins are trimers
with extensive
0021-9193/99/$04.00+0
Copyright © 1999, American Society for Microbiology. All rights reserved.
Physicochemical Evidence that Treponema
pallidum TroA Is a Zinc-Containing Metalloprotein That Lacks
Porin-Like Structure
-barrel structure, size exclusion chromatography and
circular dichroism spectroscopy revealed that TroA was a monomer and
predominantly alpha-helical. Neutron activation, atomic absorption
spectroscopy, and anomalous X-ray scattering all demonstrated that TroA
binds zinc in a 1:1 molar stoichiometric ratio. TroA does not appear to
possess structural features consistent with those of bacterial porins.
*
Corresponding author. Present address: Center for
Microbial Pathogenesis, University of Connecticut Health Center, MC
3710, School of Medicine, 263 Farmington Ave., Farmington, CT 06030. Phone: (860) 679-8129. Fax: (860) 679-8130. E-mail:
Jradolf{at}up.uchc.edu.
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