Diacylglycerol-carrying lipoprotein of hemolymph of the American cockroach: purification, characterization, and function

Abstract

A diacylglycerol-carrying lipoprotein (DGLP) was isolated from the hemolymph of adult male and female American cockroaches, Periplaneta americana. The purification procedure involved dialysis against distilled water, precipitation at low ionic concentration at pH 6.6, and separation by column chromatography on DEAE-cellulose. The final preparation was homogeneous as judged by polyacrylamide gel electrophoresis and electron microscopy. The lipoprotein comprised over 50% of the total hemolymph protein. The DGLP molecule was almost globular in shape with a diameter of approximately 160 A. The molecular weight, determined by a sedimentation-equilibrium method, was approximately 600,000. Apoprotein of DGLP consisted of two subunits, heavy chain (mol wt 250,000) and light chain (mol wt 85,000). The total lipid content of DGLP amounted to about 50%. The lipids comprised diacylglycerol (15% of total lipid), hydrocarbons (28%), cholesterol (5%), and phospholipids (43%). n-Pentacosane, 3-methylpentacosane, and 6,9-heptacosadiene were identified as major hydrocarbons. Mannose(0.9%) and glucosamine (0.3%) were associated with apoprotein of DGLP. The capacity of the purified DGLP to accept diacylglycerol from both fat body and midgut was demonstrated in vitro; thus it was suggested that the same carrier molecule served to transport this lipid from storage site and absorption site. The possible multiple role of cockroach DGLP in transporting such lipids as diacylglycerol, cholesterol, and hydrocarbon from the site of storage, absorption, or synthesis is discussed.

Keywords

• fat body
• intestine
• lipid transport
• cholesterol
• hydrocarbon