Mutant pore
The invention relates to mutant forms of lysenin. The invention also relates to analyte characterisation using the mutant forms of lysenin.
1. An apparatus comprising a transmembrane protein pore inserted into an in vitro membrane, wherein the transmembrane protein pore comprises at least one mutant lysenin monomer comprising a variant of the sequence shown set forth in SEQ ID NO: 2, wherein the variant comprises the K45T/R amino acid substitution.
2. The apparatus of claim 1 , wherein the transmembrane protein pore is a homo-oligomeric pore, and wherein each monomer of the homo-oligomeric pore is the mutant lysenin monomer.
3. The apparatus of claim 1 , wherein the apparatus comprises a hetero-oligomeric pore, and wherein at least one monomer of the hetero-oligomeric pore is the mutant lysenin monomer.
4. An apparatus for characterizing a target analyte, the apparatus comprising a plurality of mutant lysenin monomers, wherein each mutant lysenin monomer comprises a variant of the sequence set forth in SEQ ID NO: 2, wherein the variant comprises the K45T/R amino acid substitution, and wherein the plurality of mutant lysenin monomers is inserted into a plurality of in vitro membranes.
5. The apparatus of claim 4 , wherein each of the plurality of in vitro membranes comprises a homo-oligomeric pore consisting of the mutant lysenin monomers.
6. The apparatus of claim 4 , wherein each of the plurality of in vitro membranes comprises a hetero-oligomeric pore, and wherein at least one monomer of the hetero-oligomeric pore is the mutant lysenin monomer.
7. A transmembrane protein pore comprising at least one munt lysenin monomer, wherein the lysenin monomer comprises the amino acid sequence of SEQ ID NO: 2 having the K45T/R amino acid substitution.
8. The pore of claim 7 , wherein the pore is a hetero-oligomeric pore.
9. The pore of claim 7 , wherein the pore is a homo-oligomeric pore.
10. An apparatus produced by a method comprising: (i) obtaining a transmembrane protein pore comprising at least one mutant lysenin monomer, wherein the lysenin monomer comprises the amino acid sequence of SEQ ID NO: 2 having the K45T/R amino acid substitution; and (ii) contacting the pore with an in vitro membrane such that the pore is inserted into the in vitro membrane.
11. The apparatus of claim 1 , wherein the variant further comprises one or more of the following substitutions:
E94 D/Q/G/A/K/R; or
S49 L/K.
12. The apparatus of claim 4 , wherein each variant further comprises one or more of the following substitutions:
E94 D/Q/G/A/K/R; or
S49 UK.
13. The transmembrane protein pore of claim 7 , wherein the mutant lysenin monomer further comprises one or more of the following substitutions: E94 D/Q/G/A/K/R; or S49 UK.
14. The apparatus of claim 10 , wherein the lysenin monomer further comprises one or more of the following substitutions: E94 D/Q/G/A/K/R; or S49 UK.