Regulation of protein secretion by crinophagy in perfused rat liver.
We examined the secretion of three serum proteins, albumin (RSA), alpha 2 mu-globulin (alpha 2 mu G), and transferrin (Trf), in the isolated perfused liver. Within 4 h of perfusion, only 20 to 35% of previously synthesized proteins were secreted by the liver into the recirculating medium. Low temperature inhibited the secretion of alpha 2 mu G and Trf, but not RSA. The amount of RSA secreted by the liver increased twofold in the presence of leupeptin, a proteinase inhibitor, or primaquine, a weak base capable of neutralizing acidic compartments. Neither drug affected Trf secretion, while the release of alpha 2 mu G was enhanced threefold by primaquine treatment. Only 55 to 70% of the total amount of these serum proteins present in the liver at the onset of perfusion could be accounted for after 4 h of perfusion. Our evidence suggests that these losses are due to protein degradation. The degradation of RSA and alpha 2 mu G was inhibited at 15 degrees C and by both leupeptin and primaquine. Contrary, RSA degradation was not altered when livers were perfused at 20 degrees C. Morphological techniques combined with immunological probes were utilized to identify possible intracellular sites of RSA degradation. RSA and cathepsin L were colocalized to large vacuoles found near the cell periphery. Entry of RSA into these vacuoles occurred at 20 degrees C but not at 15 degrees C. Our results using perfused rat livers suggest that as much as 40% of hepatic serum proteins are degraded via fusion of secretory vesicles with lysosomes (e.g., crinophagy).
Lenk
S E
SE
Department of Anatomy and Cell Biology, University of Florida, College of Medicine, Gainesville.
Fisher
D L
DL
Dunn
W A
WA
D000510:Alpha-Globulins / Q000378:metabolism; D000818:Animals; D001798:Blood Proteins / Q000378:metabolism*; D003080:Cold Temperature; D003513:Cycloheximide / Q000494:pharmacology; D004721:Endoplasmic Reticulum / Q000378:metabolism; D006056:Golgi Apparatus / Q000378:metabolism; D007150:Immunohistochemistry; D008099:Liver / Q000187:drug effects / Q000378:metabolism*; D008247:Lysosomes / Q000378:metabolism*; D008297:Male; D008954:Models, Biological; D010477:Perfusion; D051381:Rats; D011919:Rats, Inbred Strains; D012709:Serum Albumin / Q000378:metabolism; D014168:Transferrin / Q000378:metabolism; D014617:Vacuoles / Q000378:metabolism*
D000510:Alpha-Globulins; D001798:Blood Proteins; D012709:Serum Albumin; D014168:Transferrin; C025500:alpha(2)-microglobulin; D003513:Cycloheximide