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Contenido archivado el 2024-05-27

Single Molecule Enzymology with ClyA Nanopores

Final Report Summary - SMEN (Single Molecule Enzymology with ClyA Nanopores)

Single-molecule based techniques provide the ultimate toolkit to study complex biological systems. In single-molecule approaches, molecules do not need to be synchronised as in ensemble studies, and rare and/or transient species along a reaction pathway as well as heterogeneity and disorder in a sample can be revealed. Observing and manipulating single molecules, however, is generally complicated, time consuming and subject to several technical limitations that hamper the study of single enzymes to a few selected examples.

During this ERC project, we develop a technology that uses biological nanopores to study the dynamics and activity of study single native enzymes. Our approach is simple and inexpensive, and has a timescale resolution ranging from microseconds to several hours.

Another advantage of using biological nanopores is that the output signal is an ionic current, which is easily interfaced with electronic devices. The ability of transducing an enzymatic signal into an electrical signal will allow the manufacturing of low-cost and portable sensing devices to identify biomarkers linked to the occurrence or progression of disease.

Finally, we have isolated a biological nanopore that is large enough to study double stranded nucleic acids. This nanopore will be used in practical applications, for example as a sensor for DNA mapping and fingerprinting; or in fundamental studies, for example to study protein DNA interactions.