title: Systematic investigation of the hemolymph proteome of Manduca sexta at the fifth instar larvae stage using one- and two-dimensional proteomics platforms.
abstract: Manduca sexta is an excellent insect model for studying insect physiology, including hemolymph proteins. Larvae stages of this insect are highly damaging to tobacco leaves causing a drastic decrease in crop yield. Investigation on the larval biology should help in controlling its destructive potential, thus increasing crop yields. The hemolymph is the source of its immunity to disease and environmental factors, which invariably involves protein components. To better understand the physiology of M. sexta and the protein components expressed during its life cycle, two complementary proteomics approaches, one- and two-dimensional gel electrophoresis (1-DGE and 2-DGE) in conjunction with N-terminal amino acid sequencing and liquid chromatography-mass spectrometry, were employed to analyze the fifth instar larvae hemolymph proteins. These proteomics approaches together identified 123 proteins, which constituted a total of 58 nonredundant proteins and belonged to 10 functional categories. Defense (49%), transport and metabolism (15%), storage (9%), and metamorphosis (7%) categories were highly represented accounting for 80% of the identified proteins. Besides identification of previously reported proteins, 18 novel proteins were identified, which include the lipoprotein-releasing system transmembrane protein lolC, 50S ribosomal protein L24, inducible serine protease inhibitor 1, imaginal disk growth factor, protein disulfide-isomerase-like protein ERp57, etc. The 2-DGE data were integrated to develop a 2-D gel reference map. Data obtained from 1-DGE and 2-DGE analyses are accessible through the M. sexta proteomics portal ( http://foodfunc.agr.ibaraki.ac.jp/mansehemoprot.html). Together, this study provides evidence for the presence of a large number of functionally diverse protein families in the hemolymph of M. sexta. These proteins correlate well with the fifth instar stage, the transition from larvae to pupae.
authors.affiliation: Food Function Laboratory, School of Agriculture, Ibaraki University, Ami 300-0393, 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: Misato
authors.initials: M
authors.lastname: Hirano
authors.forename: Junko
authors.initials: J
authors.lastname: Shibato
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: Karl J
authors.initials: KJ
authors.lastname: Kramer
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Kouzuma
authors.forename: Ganesh Kumar
authors.initials: GK
authors.lastname: Agrawal
authors.forename: Masami
authors.initials: M
authors.lastname: Yonekura
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D006458:Hemolymph / Q000378:metabolism*; D007814:Larva / Q000378:metabolism*; D018402:Manduca / Q000254:growth & development / Q000378:metabolism*; D008969:Molecular Sequence Data; D020543:Proteome; D040901:Proteomics; D053719:Tandem Mass Spectrometry
chemical_list: D020543:Proteome