Biosynthesis of locust lipophorin. Apolipophorins I and II originate from a common precursor.

Abstract

Biosynthesis of apolipophorins of high density lipophorin of the locust Locusta migratoria was studied in vitro. Analysis of immunoprecipitates from homogenates of in vitro labeled fat body revealed a common precursor for apolipophorin I (apoLp-I, M(r) 220,000) and apolipophorin II (apoLp-II, M(r) 72,000) with a molecular mass of approximately 280 kDa. Pulsechase experiments showed that this high molecular mass precursor is cleaved into apoLp-I and apoLp-II which subsequently are secreted as high density lipophorin from the fat body. The time required for the complete synthesis and secretion was estimated to be approximately 35 min. Both apolipophorins are glycoproteins as demonstrated by the incorporation of [3H]mannose. Treatment of [3H]mannose-labeled apolipophorin with endoglycosidase H resulted in the complete removal of the incorporated [3H]mannose. Endoglycosidase H treatment of [3H]leucine-labeled apolipophorins caused a reduction in molecular mass of approximately 3 kDa for apoLp-I and 3.5 kDa for apoLp-II, suggesting the N-linked carbohydrate content to be 1-2 and 5%, respectively. Incubation of fat body tissue in the presence of low concentrations of tunicamycin led to the synthesis and release of nonglycosylated apolipophorins.