Recognition of lactoferrin and aminopeptidase M-modified lactoferrin by the liver: involvement of the remnant receptor.
Lactoferrin inhibits the hepatic uptake of lipoprotein remnants, and we showed earlier that arginine residues of lactoferrin are involved. In this study, lactoferrin was treated with aminopeptidase-M (APM), which resulted in removal of 14 N-terminal amino acids, including 4 clustered arginines at positions 2-5 (APM-lactoferrin). After i.v. injection into rats, 125I-APM-lactoferrin was cleared within 10 min by the liver parenchymal cells (74.7% of the dose). Binding of APM-lactoferrin to isolated parenchymal liver cells was saturable with a Kd of 186 nM (750.000 sites/cell). This is in striking contrast to the binding of lactoferrin (Kd 10 microM; 20 x 10(6) sites/cell). Preinjection of rats with 20 mg of APM-lactoferrin/kg body weight reduced the liver association of beta-VLDL by 50%, whereas lactoferrin had no effect at this dose. With isolated parenchymal liver cells, APM-lactoferrin was a more effective competitor for beta-VLDL binding than native lactoferrin (50% inhibition at 0.5 mg/ml vs. 8.0 mg/ml). We conclude that the 4-arginine cluster of lactoferrin at position 2-5 involved in the massive association of lactoferrin with the parenchymal liver cell, but is not essential for the inhibition of the lipoprotein remnant uptake. The Arg/Lys sequence at position 25-30, which resembles the binding site of apoE, may mediate the high affinity binding of lactoferrin and block the binding of beta-VLDL to the remnant receptor efficiently.
Ziere
G J
GJ
Division of Biopharmaceutics, Leiden/Amsterdam Center for Drug Research, University of Leiden, Sylvius Laboratory, The Netherlands.
Kruijt
J K
JK
Bijsterbosch
M K
MK
Berkel
T J
TJ
D000626:Aminopeptidases / Q000097:blood*; D000818:Animals; D001665:Binding Sites; D004705:Endocytosis / Q000502:physiology; D007781:Lactoferrin / Q000097:blood*; D008079:Lipoproteins, VLDL / Q000097:blood*; D008099:Liver / Q000378:metabolism*; D008297:Male; D063208:Methionyl Aminopeptidases; D051381:Rats; D017208:Rats, Wistar
D008079:Lipoproteins, VLDL; D000626:Aminopeptidases; D063208:Methionyl Aminopeptidases; D007781:Lactoferrin