Methods of enhancing translocation of charged analytes through transmembrane protein pores

摘要

The invention relates to enhancing translocation of a charged analyte through a transmembrane protein pore. Translocation is enhanced by increasing the net opposing charge of the barrel or channel and/or entrance of the pore. The invention also relates to pores enhanced in accordance with the invention.

主权项

1. A method of decreasing translocation speed of a nucleic acid strand through a transmembrane beta-barrel protein pore, comprising:
(a) increasing the net positive charge of the barrel of the pore by (i) substituting one or more positively charged amino acids into the barrel of the pore or (ii) substituting one or more negatively charged amino acids into the barrel of the pore with one or more uncharged amino acids, non-polar amino acids or aromatic amino acids; and (b) passing the nucleic acid strand through the pore, wherein increasing the net positive charge decreases the translocation speed of the nucleic acid strand through the pore, and wherein said transmembrane beta-barrel protein pore is selected from the group consisting of β-toxin, α-hemolysin, leukocidin, and outer membrane porin F (OmpF).