Calcium-dependent Golgi-vesicle fusion and cathepsin B in the conversion of proalbumin into albumin in rat liver.
1. An enzyme from rat liver that converts proalbumin into albumin is described. Partial purification, inhibitor studies and the conditions for maximum activity suggest that the enzyme is cathepsin B. 2. A membrane-bound enzyme, located mainly in lysosomes, also converts proalbumin into albumin. This appears to be a membrane-bound form of cathepsin B. 3. Isolated Golgi vesicles, incubated under conditions suitable for cathepsin B, convert endogenous proalbumin into albumin. 4. This conversion in Golgi vesicles has an absolute requirement for Ca2+ at micromolar concentrations. Mg2+ does not affect or substitute for Ca2+. Both the proalbumin and the albumin formed from it are intravesicular. 5. Converting activity is enhanced by pretreatment with the known chemical fusogen, poly(ethyleneglycol). 6. Vesicles preincubated at pH above 7 in the presence of dithiothreitol show a marked fall in converting activity. This can be partially restored by incubation with native vesicles. These results suggest that vesicle fusion is a requirement for conversion of proalbumin into albumin.
D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002403:Cathepsins / Q000378:metabolism*; D004229:Dithiothreitol / Q000494:pharmacology; D006056:Golgi Apparatus / Q000187:drug effects / Q000378:metabolism; D066298:In Vitro Techniques; D008099:Liver / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D011092:Polyethylene Glycols / Q000494:pharmacology; D011228:Prealbumin / Q000378:metabolism*; D051381:Rats; D012709:Serum Albumin / Q000096:biosynthesis / Q000378:metabolism*
D011228:Prealbumin; D012709:Serum Albumin; D011092:Polyethylene Glycols; D002403:Cathepsins; D002118:Calcium; D004229:Dithiothreitol