Mutations affecting the β-sheet at the extracellular opening of the intimin β-barrel domain lead to reduced protein levels and inefficient passenger secretion.

Reductions were seen for the two mutants where the extracellular loops were removed (Int-Strep Δloop4 and Δloop5), though these results did not reach statistical significance ( P <0.15, one-way ANOVA with Dunnett’s multiple comparisons test).

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