title: Electrophoretic mapping of highly homologous keratins: a novel marker peptide approach.
abstract: Identification of the intermediate filament proteins (IFPs) in the wool proteome has formerly been hampered by limited sequence information, the high degree of IFP homology and their close proximity on 2-DE maps. This has been partially rectified by the recent acquisition of four new Type I and two Type II wool IFP sequences. Among closely migrating proteins, such as IFP clusters in a 2-DE map, proteins with higher sequence coverage will be assigned higher scores, but the identification of unique peptides in such tight clusters may distinguish these closely migrating proteins. Two approaches were adopted for the study of wool IFPs. In the first, searches were conducted for peptides known to be unique to each member of the family in each spot. In the second, MALDI imaging was employed to examine peptides bound to a PVDF membrane from a poorly resolved part of the Type I IFP region of the 2-DE map. As a result, a distinct picture has emerged of the distribution of the six Type I and four Type II IFPs across the 2-DE wool protein map.
authors: AgResearch Ltd., Christchurch, New Zealand.
authors: Santanu
authors: S
authors: Deb-Choudhury
authors: Jeffrey E
authors: JE
authors: Plowman
authors: Ancy
authors: A
authors: Thomas
authors: Gail L
authors: GL
authors: Krsinic
authors: Jolon M
authors: JM
authors: Dyer
authors: Stefan
authors: S
authors: Clerens
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D004800:Enzymes, Immobilized / Q000378:metabolism; D053530:Keratins, Type I / Q000737:chemistry* / Q000378:metabolism; D053531:Keratins, Type II / Q000737:chemistry* / Q000378:metabolism; D008969:Molecular Sequence Data; D010449:Peptide Mapping / Q000379:methods*; D011145:Polyvinyls; D034561:Sheep, Domestic; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D014935:Wool / Q000737:chemistry; D005465:Fluorocarbon Polymers
chemical_list: D004800:Enzymes, Immobilized; D053530:Keratins, Type I; D053531:Keratins, Type II; D011145:Polyvinyls; C024865:polyvinylidene fluoride; D005465:Fluorocarbon Polymers