Purification and characterization of vitellogenin from the American cockroach,Periplaneta americana |
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Institution: | 1. Institute of Marine Research, Austevoll Research Station, 5392 Storebø, Norway;2. Carolina AquaGyn, P.O. Box 12914, Raleigh, NC 27605, USA;3. INRAE, LPGP, 35000 Rennes, France |
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Abstract: | - 1.1. Vitellogenin (VG) was isolated and purified from the hemolymph of female American cockroaches.
- 2.2. The purification method used in this study comprises two steps: the first step is based on the method originally developed for purifying lipophorin from hemolymph, and the second step is the separation of VG from lipophorin by a KBr density gradient ultracentrifugation.
- 3.3. The purified VG was characterized according to molecular weight, substructure, shape and size, and lipid composition.
- 4.4. The VG molecule is almost globular in shape with the diameter of about 15.5 nm and is indistinguishable from lipophorin in shape and size.
- 5.5. The native molecular weight determined by light scattering method was 560 kDa.
- 6.6. The VG consists of four subunits with molecular weights of approximately 102, 81, 49 and 40 kDa, respectively.
- 7.7. VG is a lipoprotein and comprises 92% protein and 8% lipid.
- 8.8. Major lipid components were found to be diacylglycerol (25%) and phospholipids (71%).
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