ArticleStochastic Sensing of Nanomolar Inositol 1,4,5-Trisphosphate with an Engineered Pore

Add time:09/25/2019    Source:sciencedirect.com

The introduction of a ring of arginine residues near the constriction in the transmembrane β barrel of the staphylococcal α-hemolysin heptamer yielded a pore that could be almost completely blocked by phosphate anions at pH 7.5. Block did not occur with other oxyanions, including nitrate, sulfate, perchlorate, and citrate. Based on this finding, additional pores were engineered with high affinities for important cell signaling molecules, such as the Ca2+-mobilizing second messenger inositol 1,4,5-trisphosphate (IP3), that contain phosphate groups. One of these engineered pores, PRR-2, provides a ring of fourteen arginines that project into the lumen of the transmembrane barrel. Remarkably, PRR-2 bound IP3 with low nanomolar affinity while failing to bind another second messenger, adenosine 3′, 5′-cyclic monophosphate (cAMP). The engineered α-hemolysin pores may be useful as components of stochastic sensors for cell signaling molecules.

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