Lipid-free anchoring of thermophilic bacteriophage G20c portal adapter into solid-state nanopores
Hybrid nanopores, comprising a protein pore supported within a solid-state membrane, which combine the robust nature of solid-state membranes with the easily tunable and precise engineering of protein nanopores. In an embodiment, a lipid-free hybrid nanopore comprises a water soluble and stable, modified portal protein of the Thermus thermophilus bacteriophage G20c, electrokinetically inserted into a larger nanopore in a solid-state membrane. The hybrid pore is stable and easy to fabricate, and exhibits low peripheral leakage, allowing sensing and discrimination among different types of biomolecules.
1. A sensor comprising:
a solid-state matrix comprising a solid-state pore opening; and
a hydrophilic protein channel in a stable insertion fit within the solid-state pore opening, the hydrophilic protein channel comprising a protein nanopore channel;
wherein a protein forming at least part of the hydrophilic protein channel comprises a modification of SEQ ID NO: 1, the modification comprising one of: SEQ ID NO: 27; SEQ ID NO: 11; SEQ ID NO: 37; SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 19, SEQ ID NO: 9, SEQ ID NO: 13, SEQ ID NO: 29, SEQ ID NO: 31, SEQ ID NO: 35, SEQ ID NO: 33, SEQ ID NO: 3, SEQ ID NO: 5, and SEQ ID NO: 7.
2. The sensor of claim 1 , wherein the protein of the hydrophilic protein channel comprises SEQ ID NO: 27.
3. The sensor of claim 1 , wherein the protein of the hydrophilic protein channel comprises SEQ ID NO: 11 or SEQ ID NO: 37.
4. The sensor of claim 1 , wherein the protein of the hydrophilic protein channel comprises one of: SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 21, SEQ ID NO: 23 and SEQ ID NO: 25.
5. The sensor of claim 1 , wherein the protein of the hydrophilic protein channel comprises SEQ ID NO: 19.
6. The sensor of claim 1 , wherein the protein of the hydrophilic protein channel comprises one of: SEQ ID NO: 9 and SEQ ID NO: 13.
7. The sensor of claim 1 , wherein the protein of the hydrophilic protein channel comprises one of: SEQ ID NO: 29, SEQ ID NO: 31 and SEQ ID NO: 35.
8. The sensor of claim 1 , wherein the protein of the hydrophilic protein channel comprises SEQ ID NO: 33.
9. The sensor of claim 1 , wherein the protein of the hydrophilic protein channel comprises one of: SEQ ID NO: 3, SEQ ID NO: 5 and SEQ ID NO: 7.
10. The sensor of claim 1 , further comprising a voltage source configured to apply a voltage to an electrolyte solution on both sides of the solid-state matrix.
11. The sensor of claim 1 , wherein the solid-state matrix comprises at least one of: silicon, hafnium and nickel.
12. The sensor of claim 11 , wherein the solid-state matrix comprises at least one of: a silicon containing nitride, a silicon containing carbide and a silicon containing oxide.
13. The sensor of claim 1 , wherein the solid-state matrix comprises a thickness of less than about 30 nm.
14. The sensor of claim 1 , wherein the solid-state pore opening comprises a diameter of between about 5.4 nm and about 6 nm.
15. The sensor of claim 1 , further comprising a coating on the solid-state matrix to promote binding of the solid-state matrix to a protein forming at least part of the hydrophilic protein channel.
16. The sensor of claim 15 , wherein the coating comprises a thiol-coupling compound.
17. The sensor of claim 16 , wherein the coating comprises a maleimide compound.
18. The sensor of claim 1 , wherein a protein forming at least part of the hydrophilic protein channel comprises a modification of SEQ ID NO: 1, the modification comprising one of: SEQ ID NO: 27; SEQ ID NO: 37; SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 19, SEQ ID NO: 9, SEQ ID NO: 13, SEQ ID NO: 29, SEQ ID NO: 31, SEQ ID NO: 35, SEQ ID NO: 33, SEQ ID NO: 3, SEQ ID NO: 5, and SEQ ID NO: 7.