Purification and characterization of the common yolk protein, vitellin, from the estuarine amphipod Leptocheirus plumulosus.
Vitellin is a major yolk protein that plays a significant role in the embryonic development of crustacean embryos. This protein was rapidly purified from embryos of the estuarine amphipod, Leptocheirus plumulosus, by subjecting the crude protein homogenate to high affinity column chromatography. SDS-PAGE revealed a single band with an approximate molecular weight of 200,000 daltons. Vitellin was characterized by SDS-PAGE techniques and amino acid composition analysis. L. plumulosus vitellin is a lipoglycophosphoprotein with serine, glutamic acid/glutamine, alanine, and aspartic acid/asparagine accounting for almost 66% of all amino acid residues. Polyclonal antibodies were raised against L. plumulosus vitellin and antibody reactivity was verified by dot-blotting and immuno-fluorescence confocal microscopy. These antibodies are specific for purified vitellin and show little cross-reactivity with other embryonic proteins.
Volz
David C
DC
Department of Environmental Health Sciences, Norman J. Arnold School of Public Health, University of South Carolina, Columbia 29208, USA.
Kawaguchi
Tomohiro
T
Chandler
G Thomas
GT
D000596:Amino Acids / Q000032:analysis; D033304:Amphipoda / Q000737:chemistry*; D000818:Animals; D002846:Chromatography, Affinity / Q000379:methods; D004527:Egg Proteins / Q000737:chemistry / Q000302:isolation & purification*; D005260:Female; D005455:Fluorescent Antibody Technique; D005456:Fluorescent Dyes; D007150:Immunohistochemistry
D000596:Amino Acids; D004527:Egg Proteins; D005456:Fluorescent Dyes