Transglutaminase-catalyzed incorporation of dansyl cadaverine and dansyl-aminohexyl QQIV into MAP-rich tubulin

Solutions of MAP-rich tubulin (1 mg/mL) in 0.1 mL of 20 mM TrisCl pH 8.5 containing 2.5 mM calcium chloride, 1 mM dithiothreitol and 1 mM dansyl cadaverine or 1 mM dansyl-aminohexyl QQIV were incubated with 3 μL of 3 U/μL human transglutaminase for 16 h at 37˚C. Proteins in the reaction mixture were separated on SDS gels and stained with Coomassie blue. Seven bands were cut from the gel at about 15, 30, 50, 90, and 150 kDa plus the top of the separating gel and the top of the stacking gel. Proteins in gel slices were digested with trypsin and prepared for liquid chromatography tandem mass spectrometry as described [28].

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