Abstract
Conventional protein sequencing methods and recently reported
theoretical and experimental single molecule methods are based on analog
measurements. Here a digital method for use in the bulk or with a small
number of molecules is introduced. It is based on the superspecificity
property of transfer RNAs (tRNAs) and requires only one binary
measurement per residue/amino acid (AA). When the terminal residues of
20 copies of a protein molecule are cleaved and each brought together
with a different tRNA in one of 20 different cavities, tRNA-AA binding
occurs in exactly one cavity. The bound AA is chemically separated from
the tRNA into a channel where its presence can be detected electrically
or optically. The result is a binary high output from the cognate
channel that is independent of the AA, the other 19 outputs are low, and
the AA’s identity is given by the position of the high bit in the 20-bit
output. One way to detect the freed AA is to translocate it through a
nanopore in an electrolytic cell under electrophoresis. A feasibility
and accuracy analysis and related computations and simulation results
are presented.