The Cell Wall Teichuronic Acid Synthetase (TUAS) Is an Enzyme Complex Located in the Cytoplasmic Membrane of Micrococcus luteus
Figure 3
(a) M. luteus TUAS analyzed with native gradient PAGE. Partially purified TUAS was subjected to electrophoresis on 3–12% native gradient polyacrylamide gels (left, lane 2). Protein bands were visualized by staining with Coomassie blue. The molecular-mass standards of 545, 272, 132, and 66 kDa (top to bottom) are shown (left, lane 1). The TUAS activity is indicated with an arrow. (b) SDS-PAGE of M. luteus TUAS. Protein bands separated by 9.5% SDS-PAGE were visualized by silver staining. Lane 1: Molecular-mass standards of 97.4, 66.2, 42.7, and 31 kDa, respectively (top to bottom). Lane 2: Detergent extraction of membrane fraction. Lane 3: Protein after DEAE-cellulose column chromatography. Lane 4: TUAS purified to near homogeneity by isoelectric precipitation (pH 5.0) after DEAE-cellulose, gel filtration and adsorbent column chromatography. TUAS has two subunits A and B, upper subunit (A) is equivalent to 54 kDa and the lower subunit (B) to 52.5 kDa.
