Regions located in both the N-lobe and C-lobe of human lactoferrin participate in the binding interaction with bacterial lactoferrin receptors.
As a first step in localizing the regions of human lactoferrin involved in binding to bacterial lactoferrin receptors, N-lobe and C-lobe fragments were assessed for binding to receptors on Neisseria meningitidis, Neisseria gonorrhoeae and Moraxella (Branhamella) catarrhalis. Preparations of N-lobe and C-lobe were obtained by tryptic digestion of iron-loaded human lactoferrin followed by separation of the two lobes by gel exclusion chromatography in 10% acetic acid. Solid phase binding studies demonstrated that the isolated C- and N-lobe preparations were capable of binding to membranes from iron-deficient N. meningitidis, N. gonorrhoeae and M. catarrhalis. The binding of the individual C- and N-lobes was confirmed by an analytical SDS-PAGE binding method in which the membrane-associated polypeptides were identified by prior biotinylation and subsequent binding of labelled streptavidin. This contrasts with bacterial transferrin receptors, which only bind to C-lobe fragment of human transferrin, indicating that the bacterial lactoferrin and transferrin receptors differ in their interaction with their respective glycoprotein ligands and may differ in the mechanism of iron removal.
Yu
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RH
Department of Microbiology and Infectious Diseases, University of Calgary, Alberta, Canada.
Schryvers
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AB
D001710:Biotin; D002462:Cell Membrane / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D007501:Iron / Q000378:metabolism; D007781:Lactoferrin / Q000378:metabolism*; D001936:Moraxella catarrhalis / Q000378:metabolism*; D009344:Neisseria gonorrhoeae / Q000378:metabolism*; D009345:Neisseria meningitidis / Q000378:metabolism*; D010446:Peptide Fragments / Q000302:isolation & purification / Q000378:metabolism*; D011956:Receptors, Cell Surface / Q000302:isolation & purification / Q000378:metabolism*; D014357:Trypsin
D010446:Peptide Fragments; D011956:Receptors, Cell Surface; C038340:lactoferrin receptors; D001710:Biotin; D007501:Iron; D007781:Lactoferrin; D014357:Trypsin