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                <term xml:lang="en">calorimetry</term>
                <term xml:lang="en">intrabody</term>
                <term xml:lang="en">ribosome display</term>
                <term xml:lang="en">Sac7d</term>
                <term xml:lang="en">type II secretion system</term>
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              <p>We engineered a class of proteins that binds selected polypeptides with high specificity and affinity. Use of the protein scaffold of Sac7d, belonging to a protein family that binds various ligands, overcomes limitations inherent in the use of antibodies as intracellular inhibitors: it lacks disulfide bridges, is small and stable, and can be produced in large amounts. An in vitro combinatorial/selection approach generated specific, high-affinity (up to 140 pM) binders against bacterial outer membrane secretin PulD. When exported to the Escherichia coli periplasm, they inhibited PulD oligomerization, thereby blocking the type II secretion pathway of which PulD is part. Thus, high-affinity inhibitors of protein function can be derived from Sac7d and can be exported to, and function in, a cell compartment other than that in which they are produced.</p>
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              <org type="consortium">We thank Patrick England and Sylviane Hoos (PF5, Institut Pasteur) for help with BIAcore, Andreas Plückthun (University of Zurich, Zurich, Switzerland) for the gift of pRDV, and Nicolas Bayan for helpful advice.</org>
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