Purification and characterization of biliverdin-binding vitellogenin from the hemolymph of the common cutworm, Spodoptera litura.
Biliverdin-binding vitellogenin (Vg) was purified from adult female hemolymph of the common cutworm, Spodoptera litura, by using gel filtration and ion exchange chromatographies. The molecular mass of the protein was 490 kDa and it was composed of two 188-kDa subunits. Three internal amino acid sequences obtained by digestion of the protein with lysylendopeptidase showed high similarity to those of Bombyx mori Vg, supporting the purified blue protein to be vitellogenin. latroscan analyses demonstrated the presence of biliverdin in Vg that occupied 2.4% of total lipid components. Among the lipids of Vg (9.5 micrograms total lipids per 100 micrograms protein), diacylglycerol was the most predominant, followed by phospholipid, hydrocarbons, and then triacylglycerol, while in biliverdin-binding proteins (BPs) purified from larval hemolymph (3.1 micrograms total lipids per 100 micrograms protein), phospholipid was the most abundant lipid followed by diacylglycerol; hydrocarbons and triacylglycerol were minor components. Vg was first detected in the hemolymph of female pupae one day before eclosion, but injection of 5 micrograms of methoprene into a 3-day-old pupa induced Vg in the hemolymph 4 days earlier than in the control. Methoprene also induced a faster decline in BP-A and BP-B titers in the hemolymph with a corresponding increase of the Vg titer. These results suggest that juvenile hormone (JH) induces not only vitellogenesis but also the uptake of these proteins by stimulating the metamorphosis of fat body during the pupal stage.
Maruta
Kousei
K
Department of Applied Biological Sciences, Saga University, Saga, Japan.
Yoshiga
Toyoshi
T
Katagiri
Chihiro
C
Ochiai
Masanori
M
Tojo
Sumio
S
D000595:Amino Acid Sequence; D000818:Animals; D001664:Biliverdine / Q000378:metabolism*; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006458:Hemolymph / Q000737:chemistry / Q000378:metabolism*; D015151:Immunoblotting; D005779:Immunodiffusion; D019476:Insect Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism; D008297:Male; D008726:Methoprene / Q000494:pharmacology; D008969:Molecular Sequence Data; D008970:Molecular Weight; D017386:Sequence Homology, Amino Acid; D018411:Spodoptera / Q000737:chemistry / Q000378:metabolism*; D014819:Vitellogenins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism
D019476:Insect Proteins; D014819:Vitellogenins; D008726:Methoprene; D001664:Biliverdine