FIGURE 4.
PI-PLC release of MT6-MMP Cys mutants. COS-1 cells were transiently transfected to express with type (WT) MT6-MMP (lanes 1 and 1′) or Cys MT6-MMP mutants (lanes 2–5 and 2′–5′). Twenty-four hours later the cells were treated with 0.5 ml of PI-PLC (0.3 units/well) in phosphate-buffered saline for 30 min on ice. The supernatant was concentrated and mixed with Laemmli SDS-sample buffer with (+) or without (-) β-ME and resolved by 7.5% SDS-PAGE followed by immunoblot analysis with mAb MAB1142. MT6D, MT6-MMP dimeric form; MT6M, MT6-MMP monomeric form.