Characterization of receptors for alpha 2-macroglobulin-trypsin complex in rat hepatocytes.
High-affinity receptors for alpha 2-macroglobulin-trypsin complex were demonstrated in rat hepatocytes at 4 degrees C. The dissociation rate constant for the labelled complex was very small at low receptor occupancies, approx. 4 X 10(-4) min-1. Dissociation was biphasic at high receptor occupancies with a rate constant for the rapid phase of about 2 X 10(-2) min-1. At near-equilibrium, half of the receptors were saturated at a complex concentration of 150 pM, and the Scatchard plot was concave upwards. Thus, the binding shows complex kinetics with the probable involvement of negative cooperativity. Binding of the labelled complex was not influenced by galactose, mannose, mannose phosphate or fucoidin, whereas it was abolished in the absence of extracellular Ca2+ and inhibited by bacitracin. Approx. 70% of the labelled complex bound at 4 degrees C was rapidly internalized (kint about 3 X 10(-1) min-1) after being warmed to 37 degrees C. Radioactivity released from the cells at 37 degrees C comprised intact labelled complex and iodide. The complex was initially released at a rapid rate (k-1 about 1 X 10(-1) min-1) from about 25% of the cell-bound pool. This probably represents dissociation from the receptors. A slow phase of release followed, so that half of the bound pool was finally released as intact complex. Iodide release followed a sigmoidal curve after a 20 min lag period. Thus, specific high-affinity receptors mediate the internalization and eventual degradation of alpha 2-macroglobulin-proteinase complex into hepatocytes.
Gliemann
J
J
Davidsen
O
O
D000818:Animals; D001414:Bacitracin / Q000494:pharmacology; D002118:Calcium / Q000502:physiology; D004705:Endocytosis / Q000187:drug effects; D066298:In Vitro Techniques; D008099:Liver / Q000378:metabolism*; D026503:Low Density Lipoprotein Receptor-Related Protein-1; D008297:Male; D051381:Rats; D011971:Receptors, Immunologic / Q000378:metabolism*; D013997:Time Factors; D014357:Trypsin / Q000378:metabolism*; D000511:alpha-Macroglobulins / Q000378:metabolism*
D026503:Low Density Lipoprotein Receptor-Related Protein-1; D011971:Receptors, Immunologic; D000511:alpha-Macroglobulins; D001414:Bacitracin; D014357:Trypsin; D002118:Calcium