title: In-gel protein N- and C-termini identification and its application for transgenic protein characterization.
abstract: A new method for the determination of N- and C-termini of a protein isolated in a polyacrylamide gel is introduced. In-gel partial protein hydrolysis by hydrochloric acid is used to generate N- and C-terminal peptides for identification. This new method is complementary to existing techniques. The application of the in-gel protein termini identification technique to the characterization of the transgenic protein diacylglycerol acyltransferase (UrDGAT2A) purified from soybean seeds is also reported here. Both N- and C-termini of UrDGAT2A were successfully identified and the N-terminus was found to be blocked by acetylation. The analysis results of UrDGAT2A and two commercial proteins (bovine serum albumin (BSA) and alcohol dehydrogenase) are used to demonstrate the effectiveness of the method in identifying actual N- and C-termini, terminal truncation and blocking.
authors.affiliation: Monsanto Company, 800 N. Lindbergh Blvd., St. Louis, MO 63167, USA. bosong.xiang@monsanto.com
authors.forename: Bosong
authors.initials: B
authors.lastname: Xiang
authors.forename: Susan
authors.initials: S
authors.lastname: Macisaac
authors.forename: Kathryn
authors.initials: K
authors.lastname: Lardizabal
authors.forename: Bin
authors.initials: B
authors.lastname: Li
mesh_terms: D020816:Amino Acid Motifs; D000595:Amino Acid Sequence; D000818:Animals; D051048:Diacylglycerol O-Acyltransferase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D006868:Hydrolysis; D008969:Molecular Sequence Data; D010449:Peptide Mapping / Q000379:methods*; D010455:Peptides / Q000737:chemistry; D010940:Plant Proteins / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D012710:Serum Albumin, Bovine / Q000737:chemistry*; D013025:Glycine max / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
chemical_list: D010455:Peptides; D010940:Plant Proteins; D012710:Serum Albumin, Bovine; D051048:Diacylglycerol O-Acyltransferase