title: Alkylation of galectin-1 with iodoacetamide and mass spectrometric mapping of the sites of incorporation.
abstract: Galectins can display unique sensitivity to oxidative changes that result in significant conformational alterations that prevent carbohydrate recognition. While a variety of approaches can be utilized to prevent galectin oxidation, several of these require inclusion of reducing agents that not only prevent galectins from undergoing oxidative inactivation, but can also interfere with normal redox potentials required for fundamental cellular processes. To overcome limitations associated with placing cells in an artificial reducing environment, cysteine residues on galectins can be directly alkylated with iodoacetamide to form a stable thioether adduct that is resistant to further modification. Iodoacetamide alkylated galectin remains stable over prolonged periods of time and retains the carbohydrate binding and biological activities of the native protein. As a result, this approach allows examination of the biological roles of a stabilized form of galectin-1 without introducing the confounding variables that can occur when typical soluble reducing agents are employed.
authors.affiliation: Center for Transfusion and Cellular Therapies, Department of Pathology and Laboratory Medicine, Emory University School of Medicine, Atlanta, GA, USA.
authors.forename: Sean R
authors.initials: SR
authors.lastname: Stowell
authors.forename: Connie M
authors.initials: CM
authors.lastname: Arthur
authors.forename: Richard D
authors.initials: RD
authors.lastname: Cummings
authors.forename: Christa L
authors.initials: CL
authors.lastname: Feasley
mesh_terms: D000478:Alkylation; D001665:Binding Sites; D050260:Carbohydrate Metabolism; D002851:Chromatography, High Pressure Liquid; D037483:Galectin 1 / Q000737:chemistry* / Q000378:metabolism*; D007460:Iodoacetamide / Q000378:metabolism*; D013058:Mass Spectrometry / Q000379:methods*; D010084:Oxidation-Reduction; D059748:Proteolysis; D013223:Statistics as Topic; D013438:Sulfhydryl Compounds / Q000378:metabolism
chemical_list: D037483:Galectin 1; D013438:Sulfhydryl Compounds; D007460:Iodoacetamide