Determination of the half-life of lipoprotein lipase in cultured granulosa cells.
The incorporation of [3H]leucine into granulosa lipoprotein lipase was determined by an immunoadsorbent method: a sensitive, specific, and quantitative technique using antiserum to avian adipose lipoprotein lipase coupled to Sepharose 4B as the immunoadsorbent. The sole 3H-labeled protein bound to the immunoadsorbent was shown to be lipoprotein lipase by: (1) sodium dodecyl sulfate polyacrylamide gel electrophoresis, (2) gel filtration on Sephadex G-100, and (3) isoelectric focusing in 8 M urea of the dissociated protein. As measured by a specific radioimmunoassay, all but 4.5% of total enzyme protein, from 1.45 to 380 ng avian lipase, bound to 200 microliters immunoadsorbent. This immunoadsorbent has a quantitative binding capacity (96.5%) over a 200-fold range of enzyme concentration. Using this method, the half-life of lipoprotein lipase under steady-state conditions in cultured granulosa cells was found to be 11.26 h.
D000818:Animals; D002478:Cells, Cultured; D002645:Chickens; D005260:Female; D006107:Granulosa Cells / Q000201:enzymology*; D006207:Half-Life; D007163:Immunosorbent Techniques; D007930:Leucine; D008071:Lipoprotein Lipase / Q000378:metabolism*
D008071:Lipoprotein Lipase; D007930:Leucine