Purification and characterization of honey bee vitellogenin

Diana E. Wheeler, John K. Kawooya

Research output: Contribution to journalArticle

72 Scopus citations

Abstract

A protocol has been developed for the purification of vitellogenin from the honey bee, Apis mellifera. Purification allows for the first characterization of a vitellogenin from the large order Hymenoptera. Hymenopteran vitellogenins are unusual among insect vitellogenins in that they contain only one type of apoprotein. The honey bee vitellogenin was isolated from hemolymph of honey bee queens by a combination of density gradient ultracentrifugation, ion‐exchange chromatography, and affinity chromatography. The native vitellogenin particle is a very high density glycolipoprotein containing approximately 91% protein, 7% lipid, and 2% carbohydrate. Phospholipid and diacylglycerol are the major lipid components. The equilibrium density (1.28 g/ml) is the same as that for Manduca sexta vitellogenin, which contains a much higher proportion of lipid. The covalently bound carbohydrate moiety of the particle is high in mannose. The amino acid composition of vitellogenin is similar to those of vitellogenins from other insect species. The N‐terminal amino acid sequence of the apoprotein was determined, the first such sequence for any insect vitellogenin. When analyzed by sodium dodecyl sulfate (SDS)‐gel electrophoresis A. mellifera vitellogenin resolved into a single band with an apparent Mr of 180,000. Gel filtration under reducing and native conditions yielded estimated Mr values of about 300,000.

Original languageEnglish (US)
Pages (from-to)253-267
Number of pages15
JournalArchives of insect biochemistry and physiology
Volume14
Issue number4
DOIs
StatePublished - 1990

Keywords

  • Apis
  • Hymenoptera
  • lipoprotein
  • oogenesis
  • social insects

ASJC Scopus subject areas

  • Physiology
  • Biochemistry
  • Insect Science

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