Identification and Analysis of a Ca2+-Dependent Lactoferrin Receptor in Rat Liver

Lactoferrin Binds to the Asialoglycoprotein Receptor in a Galactose-Independent Manner
  • Douglas D. McAbee
  • David J. Bennatt
  • Yuan Yuan Ling
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 443)


We identified a 45 kDa Ca2+-dependent Lf binding protein on rat hepatocytes. Dithiobis(sulfosuccimidylproprionate) (DTSSP)-crosslinked 125I-Lf to a 45 kDa adduct in a Ca2+-dependent manner on intact cells. The 125I-labeledcrosslinked complexes were absent when either surface-bound 125I-Lf was stripped prior to crosslinking or an excess of unlabeled Lf was included in the DTSSP reaction. Triton X-100 extracts of hepatocyte membrane ghosts were chromatographed on Lf-agarose, and a 45 kDa polypeptide (p45) was eluted by EGTA. Anti-p45 sera blocked vigorously 125I-Lf endocytosis to intact rat hepatocytes, confirming that p45 functions as the Ca2+-dependent Lf receptor on hepatocytes. Two tryptic fragments of p45 showed 100% identity with internal sequences (Leu121 →Lys 126 and Phe198-Lys220) of the major subunit (RHL- l) of the rat asialoglycoprotein receptor. Antisera against p45 and RHL-1 crossreacted equally well with each protein, and asialoorosomucoid blocked the binding of 125I-Lf to hepatocytes. We did not detect the minor subunits (RHL2/3) of the rat asialoglycoprotein receptor in p45 preparations from Triton X-100-extracts of hepatocytes, and 125I-Lf bound to immobilized RHL-1 but not to RHL-2/3. Exoglycosidases were used to remove terminally-exposed NeuNAc and α-and β-Gal from bovine Lf glycans, and lectin blotting confirmed that glycosidase-treated Lfs lacked detectable terminal Gal. Unexpectedly, deglycosylated Lf exhibited no loss in its ability to compete with unmodified Lf for binding to isolated hepatocytes. Moreover, β-lactose but not sucrose competed vigorously for 125I-Lf endocytosis by hepatocytes, indicating that Lf binds at or near the carbohydrate-recognition domain of RHL-l. We conclude that RHL-1 is the Ca2+-dependent Lf receptor on hepatocytes and that it binds Lf in a Gal-independent manner.


Asialoglycoprotein Receptor Major Subunit Lectin Blotting Unbind Ligand Share Amino Acid Sequence 
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Copyright information

© Springer Science+Business Media New York 1998

Authors and Affiliations

  • Douglas D. McAbee
    • 1
  • David J. Bennatt
    • 1
  • Yuan Yuan Ling
    • 1
  1. 1.Department of Biological SciencesUniversity of Notre DameNotre DameIndiana

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