title: Extraction and identification of membrane proteins from black widow spider eggs.
abstract: The eggs of oviparous animals are storehouses of maternal proteins required for embryonic development. Identification and molecular characterization of such proteins will provide much insight into the regulation of embryonic development. We previously analyzed soluble proteins in the eggs of the black widow spider (Latrodectus tredecimguttatus), and report here on the extraction and mass spectrometric identification of the egg membrane proteins. Comparison of different lysis solutions indicated that the highest extraction of the membrane proteins was achieved with 3%-4% sodium laurate in 40 mmol/L Tris-HCl buffer containing 4% CHAPS and 2% DTT (pH 7.4). SDS-PAGE combined with nLC-MS/MS identified 39 proteins with membrane-localization annotation, including those with structural, catalytic, and regulatory activities. Nearly half of the identified membrane proteins were metabolic enzymes involved in various cellular processes, particularly energy metabolism and biosynthesis, suggesting that relevant metabolic processes were active during the embryonic development of the eggs. Several identified cell membrane proteins were involved in the special structure formation and function of the egg cell membranes. The present proteomic analysis of the egg membrane proteins provides new insight into the molecular mechanisms of spider embryonic development.
authors.affiliation: Key Laboratory of Protein Chemistry and Developmental Biology of Ministry of Education of China, Hunan Normal University, Changsha 410081, China.
authors.forename: Si-Ling
authors.initials: SL
authors.lastname: Fu
authors.affiliation: Molecular Sciences and Biomedicine Laboratory, State Key Laboratory for Chemo/Biosensing and Chemometrics, College of Biology, Collaborative Innovation Center for Chemistry and Molecular Medicine, Hunan University, Changsha 410082, China.
authors.forename: Jiang-Lin
authors.initials: JL
authors.lastname: Li
authors.affiliation: Key Laboratory of Protein Chemistry and Developmental Biology of Ministry of Education of China, Hunan Normal University, Changsha 410081, China.
authors.forename: Jia
authors.initials: J
authors.lastname: Chen
authors.affiliation: Key Laboratory of Protein Chemistry and Developmental Biology of Ministry of Education of China, Hunan Normal University, Changsha 410081, China.
authors.forename: Qiu-Ting
authors.initials: QT
authors.lastname: Wang
authors.affiliation: Key Laboratory of Protein Chemistry and Developmental Biology of Ministry of Education of China, Hunan Normal University, Changsha 410081, China.
authors.forename: Jian-Jun
authors.initials: JJ
authors.lastname: Li
authors.affiliation: Key Laboratory of Protein Chemistry and Developmental Biology of Ministry of Education of China, Hunan Normal University, Changsha 410081, China.wang_xianchun@263.net.
authors.forename: Xian-Chun
authors.initials: XC
authors.lastname: Wang
mesh_terms: D000818:Animals; D001740:Black Widow Spider / Q000737:chemistry* / Q000196:embryology; D008565:Membrane Proteins / Q000032:analysis* / Q000302:isolation & purification; D010063:Ovum / Q000737:chemistry*; D040901:Proteomics
chemical_list: D008565:Membrane Proteins
keywords: Egg; Extraction; Identification; Latrodectus tredecimguttatus; Membrane protein