The binding of lactoferrin to glycosaminoglycans on enterocyte-like HT29-18-C1 cells is mediated through basic residues located in the N-terminus.
Although lactoferrins (Lfs) isolated from milk of various mammals exhibit a close structural relationship, they show species-specific binding to cells. To define the specificity of recognition of human (hLf), bovine (bLf) and murine (mLf) lactoferrin by human intestinal cells, we analysed the binding of the three proteins to a subclone derived from human carcinoma cell line HT29. We observed that hLf and bLf interact with two types of binding sites (K(d): 63+/-22 nM; 0.7+/-0.2 microM) while mLf was recognized only by the lowest affinity binding sites with a lower number of binding sites. Using N-terminal deleted human Lf variants, we found that the sequence G(1)RRRR(5) is mainly responsible for the interactions with HT29 cells. Lactoferrin-binding sites on the surface of HT29 cells were further identified as heparan sulphate and chondroitin sulphate glycosaminoglycans. We conclude that the presence of the sequence A(1)PRK(4) in bLf and K(1)ATT(4) in mLf provides an insight into why the interaction of bLf with cell membrane-associated glycosaminoglycans is similar to that of hLf and why binding of these lactoferrin species differs from that of murine Lf.
El Yazidi-Belkoura
I
I
Laboratoire de Chimie Biologique, Université des Sciences et Technologies de Lille, Unité Mixte de Recherche du Centre National de la Recherche Scientifique, Villeneuve d' Ascq, France.
Legrand
D
D
Nuijens
J
J
Slomianny
M C
MC
van Berkel
P
P
Spik
G
G
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001665:Binding Sites; D001667:Binding, Competitive; D002417:Cattle; D002462:Cell Membrane / Q000737:chemistry / Q000378:metabolism; D002809:Chondroitin Sulfates / Q000737:chemistry; D020895:Enterocytes / Q000378:metabolism*; D006025:Glycosaminoglycans / Q000737:chemistry / Q000378:metabolism*; D019073:HT29 Cells; D006497:Heparan Sulfate / Q000737:chemistry; D006801:Humans; D007781:Lactoferrin / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008969:Molecular Sequence Data; D016415:Sequence Alignment; D013045:Species Specificity
D002809:Chondroitin Sulfates; D006025:Glycosaminoglycans; D006497:Heparan Sulfate; D007781:Lactoferrin