title: A simple method for determination of intramolecular amino acid sequence of unpurified protein. Application to human serum protein adsorbed by silica particles.
abstract: Silica particles adsorbed several kinds of human serum proteins, especially 23 kDa molecular weight protein. After SDS-PAGE of adsorbed serum proteins, gel pieces containing 23 kDa protein was cut out and set in slot of stacking gel in second SDS-PAGE following overlay of Staphylococcus aureus V8 protease. After electrophoresis, gel was subjected to electroblotting onto polyvinylidene difluoride membrane. Both bands of dye-stained 23 kDa and the peptide were cut out from membrane and analyzed for amino acid sequence. Obtained sequences agreed well with amino terminal and intramolecular sequences of human HDL-apolipoprotein, A-I.
authors.affiliation: Department of Immunology, School of Medicine, University of Occupational and Environmental Health, Kitakyushu, Japan.
authors.forename: H
authors.initials: H
authors.lastname: Nishino
mesh_terms: D000595:Amino Acid Sequence; D016632:Apolipoprotein A-I / Q000737:chemistry*; D001798:Blood Proteins / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D008969:Molecular Sequence Data; D012697:Serine Endopeptidases / Q000378:metabolism; D012822:Silicon Dioxide / Q000737:chemistry; D013194:Staining and Labeling
chemical_list: D016632:Apolipoprotein A-I; D001798:Blood Proteins; D012822:Silicon Dioxide; D012697:Serine Endopeptidases; C028344:glutamyl endopeptidase