Fig. 5.

Cwp13 cleaves SlpA but does not complement the cwp84 mutant. C. difficile cwp84 mutant harboring different plasmids was grown in BHI broth supplemented with thiamphenicol (15 μg/ml) till late exponential phase (OD₆₀₀ = 0.5). Cell wall extracts and culture supernatants were prepared and analyzed by Coomassie blue staining (top) and Western blotting with antibody against the LMW SLP (middle and bottom). The bottom panel shows the 34-kDa LMW SLP species on an overexposed Western blot. Plasmids: –, pMTL960; 84, pCwp84_WT; 84*, pCwp84_C116A; 13, pCwp13_WT; 13*, pCwp13_C109A. Open arrow, 77-kDa Cwp84; solid arrow, 78-kDa Cwp13_C109A processed by endogenous Cwp13; ◀, 47-kDa fragment of SlpA.