Purification and properties of a predominantly female-specific protein from the hemolymph of the larva of the tobacco hornworm, Manduca sexta.
A major serum protein was isolated from the hemolymph of larvae of the female tobacco hornworm, Manduca sexta, just prior to metamorphosis. After 3 or 4 days, this predominantly female-specific protein is rapidly cleared from the hemolymph and taken up and stored by the fat body. This larval serum protein was purified by density gradient ultracentrifugation, gel permeation, and ion-exchange chromatography. The purified protein exhibits a single band on native gel electrophoresis and sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Chemical cross-linking with dimethylsuberimidate indicates a hexameric subunit arrangement for the native protein. The amino acid composition, relatively rich in methionine but poor in cysteine, was used to calculate a v = 0.75 cm3/g. Analytical ultracentrifugation experiments yielded S020,w = 16.9 S and D020,w = 3.23 X 10(-7) cm2/s. From these values Mr = 510,000, f/f0 = 1.22, and Stokes radius = 66.3 A were calculated. Immunoblotting experiments with anti-larval serum protein serum indicate a cross-reactivity with storage protein-1 of Bombyx mori. The amino acid composition and immunological data suggest that larval serum protein may be an example of a class of insect storage proteins distinct from the arylphorins, which are characterized by high content of aromatic amino acids.
D000596:Amino Acids / Q000032:analysis; D000818:Animals; D002241:Carbohydrates / Q000032:analysis; D029721:Drosophila Proteins; D005216:Fat Body / Q000378:metabolism; D005260:Female; D006458:Hemolymph / Q000032:analysis*; D007122:Immunoelectrophoresis; D007163:Immunosorbent Techniques; D007301:Insect Hormones / Q000302:isolation & purification*; D007814:Larva / Q000032:analysis; D007915:Lepidoptera / Q000032:analysis*; D046911:Macromolecular Substances; D014461:Ultracentrifugation
D000596:Amino Acids; D002241:Carbohydrates; D029721:Drosophila Proteins; D007301:Insect Hormones; D046911:Macromolecular Substances; C034648:larval serum protein, Drosophila