Skip to main content
. 2010 Jun 7;285(32):24825–24833. doi: 10.1074/jbc.M110.125872

FIGURE 4.

FIGURE 4.

Expression and characterization of recombinant UXNAcS. A, SDS-PAGE of total soluble protein isolated from E. coli cells expressing UXNAcS (lane 2) or control empty vector (lane 3) and of nickel column-purified fractions (lane 4, UXNAcS; lane 5, control). B, high pressure liquid chromatogram of the UXNAcS enzyme reaction. Purified recombinant UXNAcS was incubated with UDP-GlcNAcA in the presence (panel 3) or absence (panel 4) of exogenous NAD+. As a control, the corresponding column-purified protein isolated from cells expressing control empty vector was incubated with UDP-GlcNAcA and NAD+ (panel 6). Activity of total protein isolated from cell expressing recombinant UXNAcS or vector control is shown in panels 2 and 5, respectively. The reaction products were separated on a Q15 column, and the UDP-sugar peak (marked by an asterisk in panels 2 and 3) was collected separately and analyzed by MALDI-MS and NMR.