CONDUCTIVE SYNTHETIC PEPTIDES FOR MOLECULAR ELECTRONICS
In various embodiments, a synthetic peptide finding use as a molecular wire in a molecular electronic circuit comprises an alpha helical segment further comprising repeating alpha-helical motifs. The synthetic peptide may further comprise at least one specific conjugation site between the termini for attachment to a molecule such as a binding probe, and may further comprise termini having metal binding functionality such as repeats of material binding sequences. In various aspects, the synthetic peptide comprises intramolecular hydrogen bonding, salt bridges, and optionally, aromatic rings that provide for electrical conductivity through the peptide.
1 . A synthetic peptide comprising the formula:
[X 1 X 2 ] m X 3 [X 4 ] n X 3 [X 2 X 1 ] m
wherein
each X 1 independently comprises a material binding peptide comprising about 5 to about 15 amino acids, a protease cleavage sequence, or a peptide capture tag;
each X 2 independently comprises a glycine/serine {G,S} rich linker or a C1-C20 carbon chain molecular linker;
each X 3 independently comprises a covalent bond, a single amino acid, a transitional helical-promoting motif, a metal binding group, or a material binding peptide comprising about to about 15 amino acids;
each X 4 independently comprises an alpha helix motif comprising about 4 to about 40 amino acids;
each m is independently 0 to 4; and
n is 1 to 40.
2 . The synthetic peptide of claim 1 , wherein at least one instance of X 4 comprises a conjugation site.
3 . The synthetic peptide of claim 2 , wherein the conjugation site comprises cysteine, lysine, tyrosine, a biotin, an azide, or a click chemistry group.
4 . The synthetic peptide of claim 1 , wherein at least one instance of X 1 comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 98% sequence identity to any one of SEQ ID NOs: 6, 7, 8, 9, 10, 11, 16, 18 or 29.
5 . The synthetic peptide of claim 1 , wherein at least one instance of X 2 comprises glycine, serine, GS, GSG, SEQ ID NO: 24, an amino acid sequence having at least 80% sequence identity to SEQ ID NO: 25, or a C1-C20 carbon chain molecular linker.
6 . The synthetic peptide of claim 1 , wherein in any one grouping of [X 1 X 2 ] m X 3 , if m≠0, then X 3 comprises a covalent bond, a single amino acid, or a transitional helical-promoting motif.
7 . The synthetic peptide of claim 1 , wherein in any one grouping of [X 1 X 2 ] m X 3 , if m≠0, then X 3 comprises a metal binding group or a material binding peptide comprising about 5 to about amino acids.
8 . The synthetic peptide of claim 1 , wherein the metal binding group comprises C, CC, CCC, SEQ ID NO: 32, SEQ ID NO: 33, SEQ ID NO: 5, SEQ ID NO: 35, or a FLASH binding motif of the sequence CCXXCC wherein X is any amino acid.
9 . The synthetic peptide of claim 1 , wherein at least one instance of X 4 comprises an amino acid sequence having at least 80%, at least 85%, at least 90%, at least 95%, or at least 98% sequence identity to any one of SEQ ID NOs: 1, 2, 3, 4, 13, 17, 20, 22, 23, 26, 27, 28 or 31.
10 . The synthetic peptide of claim 1 comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, or at least 98% sequence identity to SEQ ID NO: 14.
11 . The synthetic peptide of claim 1 comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, or at least 98% sequence identity to SEQ ID NO: 15.
12 . The synthetic peptide of claim 1 comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, or at least 98% sequence identity to SEQ ID NO: 19.
13 . The synthetic peptide of claim 1 comprising an amino acid sequence with at least 80%, at least 85%, at least 90%, at least 95%, or at least 98% sequence identity to SEQ ID NO: 21.
14 . A molecular electronics circuit comprising:
a first electrode;
a second electrode spaced apart from the first electrode by a nanogap;
a bridging molecular wire comprising a synthetic peptide according to claim 2 , electrically connected to both the first and second electrodes to bridge the nanogap; and
a polymerase enzyme conjugated to the conjugation site,
wherein the circuit includes a conductive pathway through the synthetic peptide.
15 . A sensor comprising:
the molecular electronics circuit of claim 14 ; and
a trans-impedance amplifier is electrically connected to at least one of the first electrode and second electrode, the trans-impedance amplifier providing an output comprising a measurable electrical parameter.
16 . A CMOS chip device comprising an array of the sensors according to claim 15 .
17 . A method of sequencing a DNA molecule, comprising:
providing the sensor of claim 15 ;
initiating at least one of a voltage or a current through the circuit;
exposing the circuit to a solution containing primed single stranded DNA and/or dNTPs; and
measuring electrical signals through the circuit as the polymerase engages and extends a template,
wherein the electrical signals are processed to identify features that provide information on the underlying sequence of the DNA molecule processed by the polymerase.
18 . A molecular electronics circuit comprising:
a first electrode and a second electrode spaced apart by a nanogap;
a first synthetic peptide according to claim 1 electrically connected between the first electrode and a first site of a polymerase enzyme;
a second synthetic peptide according to claim 1 electrically connected between the second electrode and a second site of the polymerase enzyme,
wherein the circuit includes a conductive pathway through a portion of the polymerase enzyme.
19 . A sensor comprising:
the molecular electronics circuit of claim 18 ; and
a trans-impedance amplifier is electrically connected to at least one of the first electrode and second electrode, the trans-impedance amplifier providing an output comprising a measurable electrical parameter.
20 . A CMOS chip device comprising an array of the sensors according to claim 19 .
21 . A method of sequencing a DNA molecule, comprising:
providing the sensor of claim 19 ;
initiating at least one of a voltage or a current through the circuit;
exposing the circuit to a solution containing primed single stranded DNA and/or dNTPs; and
measuring electrical signals through the circuit as the polymerase engages and extends a template,
wherein the electrical signals are processed to identify features that provide information on the underlying sequence of the DNA molecule processed by the polymerase.