Purification and characterization of honey bee vitellogenin.

Diana E Wheeler, J. K. Kawooya

Research output: Contribution to journalArticle

72 Citations (Scopus)

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
StatePublished - 1990

Fingerprint

Vitellogenins
Honey
Bees
vitellogenin
honey bees
Purification
Insects
apoproteins
Apoproteins
Lipids
Apis mellifera
insects
lipids
Gels
Carbohydrates
queen honey bees
carbohydrates
Manduca
Affinity chromatography
Amino Acids

ASJC Scopus subject areas

  • Insect Science
  • Biochemistry, Genetics and Molecular Biology(all)
  • Physiology
  • Physiology (medical)

Cite this

Purification and characterization of honey bee vitellogenin. / Wheeler, Diana E; Kawooya, J. K.

In: Archives of Insect Biochemistry and Physiology, Vol. 14, No. 4, 1990, p. 253-267.

Research output: Contribution to journalArticle

@article{1be3080d1b1645b3bfe8d882f3fdc694,
title = "Purification and characterization of honey bee vitellogenin.",
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.",
author = "Wheeler, {Diana E} and Kawooya, {J. K.}",
year = "1990",
language = "English (US)",
volume = "14",
pages = "253--267",
journal = "Archives of Insect Biochemistry and Physiology",
issn = "0739-4462",
publisher = "John Wiley and Sons Ltd",
number = "4",

}

TY - JOUR

T1 - Purification and characterization of honey bee vitellogenin.

AU - Wheeler, Diana E

AU - Kawooya, J. K.

PY - 1990

Y1 - 1990

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=0025524640&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025524640&partnerID=8YFLogxK

M3 - Article

C2 - 2134180

AN - SCOPUS:0025524640

VL - 14

SP - 253

EP - 267

JO - Archives of Insect Biochemistry and Physiology

JF - Archives of Insect Biochemistry and Physiology

SN - 0739-4462

IS - 4

ER -