Cryptic domains of a 60 kDa heat shock protein of Helicobacter pylori bound to bovine lactoferrin

Hamid Reza Amini, Felipe Ascencio*, Eduardo Ruiz-Bustos, Maria J. Romero, Torkel Wadström

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

10 Scopus citations

Abstract

Bovine lactoferrin binds to a 60 kDa heat shock protein of Helicobacter pylori. Binding ability was related to human immunoglobulin G because bovine lactoferrin binding proteins were isolated by extraction of cell surface associated proteins with distilled water, applied on IgG-Sepharose and nickel sulphate chelate affinity chromatography. Binding was demonstrated by Western blot after purified protein was digested with α-chymotrypsin and incubated with peroxidase-labeled bovine lactoferrin. Binding was inhibited by bovine lactoferrin, lactose, rhamnose, galactose, and two iron-containing proteins, ferritin and haptoglobin. Helicobacter pylori binds ferritin and haptoglobin via charge or hydrophobic interactions because this binding was not inhibited by specific and various glycoproteins or carbohydrates. Carbohydrate moieties of bovine lactoferrin molecules seem to be involved in binding because glycoproteins with similar carbohydrate structures strongly inhibited binding. Scatchard plot analysis of the binding of peroxidase-labeled bovine lactoferrin to H. pylori cells yielded a k(d) 2.88 x 10-6 M. In addition, binding of H. pylori cells to bovine lactoferrin was enhanced when bacteria treated with pepsin or α-chymotrypsin after isolation from iron-restricted and iron-containing media.

Original languageEnglish
Pages (from-to)247-255
Number of pages9
JournalFEMS Immunology and Medical Microbiology
Volume16
Issue number3-4
DOIs
StatePublished - 31 Dec 1996
Externally publishedYes

Bibliographical note

Funding Information:
This study was supported by grants from the Swedish Medical Research Council (16X 047231, Medical Faculty of Lund University, and the Center for Biological Research (BIT-2). We appreciatet he assistanceo f Dr. Ellis Glazier in editing the English

Keywords

  • Helicobacter pylori
  • heat shock protein
  • lactoferrin
  • lactoferrin-binding protein

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