Multipartite and circularly permuted beta-barrel polypeptides and methods for their use
AssigneeUniversity of Washington
InventorsJason C. KLIMA, David BAKER
Life of this patent
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Abstract
Disclosed herein are β-barrel polypeptides including self-complementing multipartite β-barrel polypeptides and circularly permuted β-barrel polypeptides and methods for their design and use in mediating real-time monitoring of polypeptide-polypeptide association and dissociation events.
Claims25
A non-naturally occurring, self complementing multipartite β-barrel protein, comprising at least a first polypeptide component and a second polypeptide component, wherein the at least first polypeptide component and the second polypeptide component are not covalently linked, wherein in total the at least first polypeptide component and the second polypeptide component comprise domains X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14-X15-X16-X17-X18-X19, wherein:
- X1 comprises a capping domain;
- X2 comprises a beta strand;
- wherein a contiguous C-terminal portion of X1 and N-terminal portion of X2 comprise the amino acid sequence Z1-P-G-Z2-W, where Z1 and Z2 are any amino acid;
- X3 comprises a beta turn;
- X4 comprises a beta strand that includes an internal G residue and a P at its C-terminus;
- X5 comprises a single polar amino acid;
- X6 comprises a beta turn;
- X7 comprises a beta strand including an internal G residue;
- X8 comprises a beta turn;
- X9 comprises a beta strand including an internal P residue and 2 internal G residues;
- X10 comprises a single polar amino acid;
- X11 comprises a beta turn;
- X12 comprises a beta strand;
- X13 comprises a beta turn;
- X14 comprises a beta strand with an internal G residue;
- X15 comprises a single polar amino acid;
- X16 comprises a beta turn;
- X17 comprises a beta strand;
- X18 comprises a beta turn;
- and
- X19 comprises a beta strand;
- wherein (a) each beta strand is fully present within one polypeptide component of the at least first polypeptide component and the second polypeptide component, (b) none of the at least first polypeptide component and the second polypeptide component include each of X2, X4, X7, X9, X12, X14, X17, and X19;
- and (c) one of domains X3, X6, X8, X11, X13, X16, and X18 may be partially or wholly absent in each of the first polypeptide and the second polypeptide.
(canceled)
The self-complementing multipartite β-barrel protein of claim wherein Z1 is a hydrophobic amino acid and Z2 is a polar amino acid.
-25. (canceled)
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A β-barrel polypeptide, comprising domains X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14-X15-X16-X17-X18-X19, wherein:
- X1 comprises a capping domain;
- X2 comprises a beta strand,
- wherein a contiguous C-terminal portion of X1 and N-terminal portion of X2 comprise the amino acid sequence Z1-P-G-Z2-W, where Z1 and Z2 are any amino acid;
- X3 comprises a beta turn;
- X4 comprises a beta strand that includes an internal G residue and a P at its C terminus;
- X5 comprises a single polar amino acid;
- X6 comprises a beta turn;
- X7 comprises a beta strand including an internal G residue;
- X8 comprises a beta turn;
- X9 comprises a beta strand including an internal P residue and 2 internal G residues;
- X11.0 comprises a single polar amino acid;
- X11 comprises a beta turn;
- X12 comprises a beta strand;
- X13 comprises a beta turn;
- X14 comprises a beta strand with an internal G residue;
- X15 comprises a single polar amino acid;
- X16 comprises a beta turn;
- X17 comprises a beta strand;
- X18 comprises a beta turn;
- and
- X19 comprises a beta strand;
- wherein the last residue of the X19 domain is N-terminal to and connected to the first residue of X1 domain via an amino acid linker;
- wherein 1, 2, or 3 contiguous domains X1, X2, X3, X4, X5, X6, X7, X8, X9, X10, X11, X12, X13, X14, X15, X16, X17, X18, and X19 may be partially or wholly absent.
-67. (canceled)
Dependencies are inferred from the claim text.