Identification of Lactoferrin-Binding Proteins From Treponema Pallidum Subspecies Pallidum and Treponema Denticola

Mol Microbiol. 1994 May;12(4):613-9. doi: 10.1111/j.1365-2958.1994.tb01048.x.

Abstract

Lactoferrin-binding or -associated proteins were identified in Treponema pallidum subspecies pallidum and Treponema denticola by affinity column chromatography using human lactoferrin and detergent-solubilized, radiolabelled spirochaetes. Two discrete polypeptides of T. pallidum with masses of 45 and 40 kDa and a broad band from 29-34 kDa exhibited association with human apo- and partially ferrated lactoferrin. T. denticola produced two proteins that associated with a lactoferrin affinity matrix (50 and 35 kDa). T. pallidum and T. denticola did not associate with soluble, human transferrin in parallel experiments. Soluble human lactoferrin competed with all lactoferrin-associated proteins from T. pallidum and T. denticola in competitive-binding assays. However, the T. denticola proteins dissociated from a lactoferrin-affinity matrix in the presence of differing concentrations of unlabelled, soluble lactoferrin competitor. Treatment with phospholipase D altered migration of the diffuse 29-34 kDa band of T. pallidum suggesting that the polypeptide was lipid-modified. Each of the lactoferrin-binding proteins from T. pallidum and T. denticola reacted with pooled rabbit syphilitic antisera. The lactoferrin-binding proteins of T. pallidum reacted with human sera from patients at all stages of syphilis. In addition, a monoclonal antibody generated against the 45 kDa polypeptide of T. pallidum crossreacted with the 29-34 kDa protein.

Publication types

  • Research Support, U.S. Gov't, P.H.S.

MeSH terms

  • Antibodies, Bacterial / blood
  • Bacterial Proteins / immunology
  • Bacterial Proteins / isolation & purification
  • Bacterial Proteins / metabolism
  • Binding, Competitive
  • Carrier Proteins / immunology
  • Carrier Proteins / isolation & purification*
  • Carrier Proteins / metabolism
  • Chromatography, Affinity
  • Humans
  • In Vitro Techniques
  • Lactoferrin / metabolism*
  • Molecular Weight
  • Syphilis / immunology
  • Treponema / metabolism*
  • Treponema pallidum / metabolism*

Substances

  • Antibodies, Bacterial
  • Bacterial Proteins
  • Carrier Proteins
  • lactoferrin binding proteins, bacterial
  • Lactoferrin