Isolation and characterization of the high-density lipoproteins from the hemolymph and ovary of the penaeid shrimp Penaeus semisulcatus (de Haan): apoproteins and lipids.
The high-density lipoproteins (HDLs) found in the male and female hemolymph of Penaeus semisulcatus de Haan were isolated by NaBr (1.22 g/ml) followed by sucrose gradient (5-25%) ultracentrifugation. The male HDL contained one protein, lipoprotein 1 (LP1), composed of one 110-kDa peptide subunit. The female HDL contained two proteins: 1) the LP1 that was immunoidentical to the male LP1 and was similarly composed of one 110-kDa peptide subunit and 2) vitellogenin (Vg), reacting positively with the rabbit antiserum generated against vitellin (Vt) that was isolated from vitellogenic ovaries. Both Vg and Vt consisted mainly of three polypeptide subunits (200, 120, and 80 kDa) as revealed by denatured PAGE and Western blot. The LP1 from males or females did not react with the Vt rabbit antiserum. Similarly, Vg and Vt did not react with the rabbit antiserum prepared against LP1. Phospholipids (PL) constituted 71-76% of the total lipids in the hemolymph and HDLs of both male and female hemolymph. Cholesterol (Ch) amounted to 17-20%, and small amounts (5%) of diacylglycerols (DAG) were also carried by these HDLs. Both the PL and DAG contained highly unsaturated fatty acids (20:5 omega 3 and 22:6 omega 3) that are transported from the food or hepatopancreas to the tissues, including the vitellogenic ovaries in females. In the present study we show for the first time the separate lipid composition of female LP1 and Vg and compare them with the lipids attached to the Vt. Vg had a lower lipid content than LP1 (540 and 1089 mg/g protein, respectively). Differences were also found in the relative abundance of PL, Ch, and DAG classes in the LP1 in comparison with Vg. Furthermore, small amounts (approximately 3.8%) of triacylglycerols (TAG) were found only in the hemolymph of vitellogenic females, and they were associated with the Vg. Although Vg and Vt were composed of similar polypeptides, their lipid composition was different Vt, in contrast to Vg, carried considerable amounts of TAG (approximately 22%) and only trace amounts of DAG. The significance of the TAG in the hemolymph of vitellogenic females is not known, and the functional relationship between Vg and Vt requires future extensive studies. Lipids were not detected in hemocyanin that was purified from clotted hemolymph.
Lubzens
E
E
National Institute of Oceanography, Haifa, Israel.
Ravid
T
T
Khayat
M
M
Daube
N
N
Tietz
A
A
D000818:Animals; D015153:Blotting, Western; D002499:Centrifugation, Density Gradient; D004527:Egg Proteins / Q000032:analysis / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D005227:Fatty Acids / Q000032:analysis; D005260:Female; D006458:Hemolymph / Q000737:chemistry*; D008055:Lipids / Q000032:analysis; D008075:Lipoproteins, HDL / Q000032:analysis* / Q000302:isolation & purification*; D008297:Male; D010053:Ovary / Q000737:chemistry*; D033561:Penaeidae / Q000737:chemistry*; D012727:Sex Characteristics; D014819:Vitellogenins / Q000032:analysis / Q000302:isolation & purification
D004527:Egg Proteins; D005227:Fatty Acids; D008055:Lipids; D008075:Lipoproteins, HDL; D014819:Vitellogenins