title: Comprehensive royal jelly (RJ) proteomics using one- and two-dimensional proteomics platforms reveals novel RJ proteins and potential phospho/glycoproteins.
abstract: Royal jelly (RJ) is an exclusive food for queen honey bee (Apis mellifera L.) that is synthesized and secreted by young worker bees. RJ is also widely used in medical products, cosmetics, and as health foods. However, little is known about RJ functionality and the total protein components, although recent research is attempting to unravel the RJ proteome. We have embarked on a detailed investigation of the RJ proteome, using a modified protein extraction protocol and two complementary proteomics approaches, one- and two-dimensional gel electrophoresis (1-DGE and 2-DGE) in conjunction with tandem mass spectrometry. Simultaneously, we examined total soluble protein from RJ collected at 24, 48, and 72 h after honey bee larvae deposition twice (in two flower blooming seasons), to check differences, if any, in RJ proteome therein. Both 1- and 2-D gels stained with silver nitrate revealed similar protein profiles among these three time points. However, we observed a clear difference in two bands (ca. MW of 55 and 75 kDa) on 1-D gel between the first and the second collection of RJ. A similar difference was also observed in the 2-D gel. Except for this difference, the protein profiles were similar at the 3 time points. As the RJ from 48 (or sometimes 72) is commercially used, we selected the RJ sample at 48 h for detailed analysis with the first collection. 1-DGE identified 90 and 15 proteins from the first and second selection, respectively; in total, 47 nonredundant proteins were identified. 2-DGE identified 105 proteins comprising 14 nonredundant proteins. In total, 52 nonredundant proteins were identified in this study, and other than the major royal jelly protein family and some other previously identified proteins, 42 novel proteins were identified. Furthermore, we also report potentially post-translationally modified (phosphorylation and glycosylation) RJ proteins based on the Pro-Q diamond/emerald phosphoprotein/glycoprotein gel stains; MRJP 2p and 7p were suggested as potential phosphoproteins. The 2-DGE data were integrated to develop a 2-D gel reference map, and all data are accessible through RJ proteomics portal (http://foodfunc.agr.ibaraki.ac.jp/RJP.html).
authors.affiliation: Food Function Laboratory, School of Agriculture, Ibaraki University, Ami, Japan.
authors.forename: Takako
authors.initials: T
authors.lastname: Furusawa
authors.forename: Randeep
authors.initials: R
authors.lastname: Rakwal
authors.forename: Hyung Wook
authors.initials: HW
authors.lastname: Nam
authors.forename: Junko
authors.initials: J
authors.lastname: Shibato
authors.forename: Ganesh Kumar
authors.initials: GK
authors.lastname: Agrawal
authors.forename: Yu Sam
authors.initials: YS
authors.lastname: Kim
authors.forename: Yoko
authors.initials: Y
authors.lastname: Ogawa
authors.forename: Yasukazu
authors.initials: Y
authors.lastname: Yoshida
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Kouzuma
authors.forename: Yoshinori
authors.initials: Y
authors.lastname: Masuo
authors.forename: Masami
authors.initials: M
authors.lastname: Yonekura
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D001514:Bee Venoms / Q000032:analysis; D001516:Bees; D002853:Chromatography, Liquid; D015180:Electrophoresis, Gel, Two-Dimensional; D005227:Fatty Acids / Q000032:analysis*; D019476:Insect Proteins / Q000032:analysis*; D013058:Mass Spectrometry; D008969:Molecular Sequence Data; D020543:Proteome / Q000032:analysis*; D000099219:Royal Jelly
chemical_list: D001514:Bee Venoms; D005227:Fatty Acids; D019476:Insect Proteins; D020543:Proteome; D000099219:Royal Jelly