Sie sind hier: Startseite Aktuelles Termine-Vorträge Prof. Liviu Movileanu, Syracuse …

Prof. Liviu Movileanu, Syracuse University, New York, USA

Single-Molecule Detection of Proteins in A Complex Biofluid: Finding A Needle in A Haystack
Wann 01.07.2019
von 14:15 bis 15:15
Wo Seminarraum B, Freiburger Materialforschungszentrum (FMF)
Termin übernehmen vCal
iCal

Protein-protein interactions (PPIs) are at the heart of cell signaling. Yet, we have very limited ways to quantitate them, especially in a heterogeneous solution, such as blood serum, cell lysates, or tissue biopsies. We have manufactured a single-polypeptide chain protein sensor capable of detecting transient PPIs, one interaction at a time. This sensor is like a fishing rod with three parts: a protein receptor in the form of a hook, a hexapeptide tether, which is similar to a short, flexible line, and a signal transducer (t-FhuA), functioning like an angling rod.1, 2 The transducer is an extensive truncation of ferric hydroxamate uptake component A (FhuA), an outer-membrane, b-barrel protein of E. coli. In this way, t-FhuA forms a nanopore that facilitates the transmembrane permeation of ions, leading to a uniform current. When a free protein ligand in solution is reversibly captured by the tethered protein receptor, transient capture and release events are measured as current deflections between two open substates of the pore. Notably, these events are unambiguously discriminated in mammalian serum. Our sensor could be extended to emerging areas of molecular diagnostics or might be adapted to develop tools in high-throughput protein profiling and drug discovery.


Acknowledgements. I am thankful to my graduate student, Avi Thakur, as well as others in my laboratory and funding agency (NIH R01 GM088403 and NIH R01 GM129429).


REFERENCES
1. Thakur, A.K. & Movileanu, L. Real-time measurement of protein-protein
interactions at single-molecule resolution using a biological nanopore. Nat.
2. Biotechnol. 37, 96-101 (2019).
Kuehnel, K. Touch-and-go sensing. Nat. Methods 16, 145 (2019).

Organisation: