Plasma vitellogenin of grouper (Epinephelus malabaricus): isolation and properties.

Radiolabelled vitellogenin produced by juvenile grouper following injection of 3H-leucine, 32P-orthophosphate and estradiol-stimulation was purified by anion-exchange and gel filtration chromatography and characterized by electrophoresis and chemical analysis. The radiolabelled product was found to exist in two heterogeneous molecular weight forms by electrophoresis under nondenaturing conditions on polyacrylamide gel with M(r), of 260,000 and 525,000. It showed two protein monomers (M(r) 113,000 and 140,000) on electrophoresis under denaturing condition on a sodium dodecyl sulphate-polyacrylamide gel. The purified material contained 17.2% total lipid with 1.3% cholesterol and 2.1% triglycerides, 8-21 micrograms/mg protein total carbohydrate and 6.8 micrograms phosphate/mg protein; amino acid analysis showed a profile comparable to that found for vitellogenins isolated from the grouper, medaka, goldfish and rainbow trout. The study showed grouper vitellogenin to be a glycophospholipoprotein similar in composition to vitellogenins from other teleosts and demonstrated that vitellogenins can be induced in juveniles by injection with estradiol-17 beta.