De novo designed NTF2-like scaffolds for de novo design of enzymes and small molecule binders

Information

  • Patent Application
  • 20230295639
  • Publication Number
    20230295639
  • Date Filed
    January 17, 2023
    a year ago
  • Date Published
    September 21, 2023
    a year ago
Abstract
Polypeptide are disclosed that comprise an amino acid sequence at least 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from the group consisting of SEQ ID NO:1-1615, not including any functional domains added fused to the protein (whether N-terminal, C-terminal, or internal), and wherein the 1, 2, 3, 4, or 5 N-terminal and/or C-terminal amino acid residues may be present or absent when considering the percent identity.
Description
Claims
  • 1. A polypeptide comprising an amino acid sequence at least 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identical to the amino acid sequence selected from the group consisting of SEQ ID NO:1-1615, not including any functional domains added fused to the protein (whether N-terminal, C-terminal, or internal), and wherein the 1, 2, 3, 4, or 5 N-terminal and/or C-terminal amino acid residues may be present or absent when considering the percent identity.
  • 2. The polypeptide of claim 1, wherein 1, 2, 3, 4, or 5 N-terminal and/or C-terminal amino acid residues are absent and are not included when determining the percent identity relative to the reference amino acid sequence.
  • 3. The polypeptide of claim 1, wherein all amino acid residues in the reference sequences are included when determining the percent identity relative to the reference protein.
  • 4. The polypeptide of claim 1, wherein the polypeptide comprises the secondary structure arrangement H1-L1-H2-L2-E1-L3-E2-L4-H3-L5-E3-L6-E4-L7-E5-L8-E6, wherein “H” is a helical domain, “L” is a loop domain, and “E” is a beta strand domain.
  • 5. The polypeptide of claim 4, wherein domain H1 is 19 amino acids in length, domain H2 is 7 amino acids in length, domain H3 is 14 amino acids in length, domain E1 is 4 amino acids in length, domain E2 is 4 amino acids in length, domain E3 is 10 amino acids in length, domain E4 is 12 amino acids in length, domain E5 is 14 amino acids in length, and domain E6 is 13 amino acids in length.
  • 6. A fusion protein comprising the polypeptide of claim 1 fused to one or more functional polypeptides.
  • 7. The fusion protein of claim 6, wherein the one or more functional polypeptides are fused to the N-terminus and/or C-terminus of the polypeptide.
  • 8. The fusion protein of claim 5, wherein one or more functional polypeptides are inserted internally into the protein.
  • 9. The fusion protein of claim 8, wherein the polypeptide comprises the secondary structure arrangement H1-L1-H2-L2-E1-L3-E2-L4-H3-L5-E3-L6-E4-L7-E5-L8-E6, wherein “H” is a helical domain, “L” is a loop domain, and “E” is a beta strand domain, and wherein each of the one or more functional polypeptide inserted internally into the polypeptide is inserted into a loop domain.
  • 10. A nucleic acid encoding the protein or fusion protein of claim 1.
  • 11. An expression vector comprising the nucleic acid of claim 10 operatively linked to a suitable control sequence.
  • 12. A host cell comprising the expression vector of claim 11.
  • 13. A library comprising 5, 10, 35, 50, 100, 250, 500, 750, 1000, or more different polypeptides according to claim 1.
  • 14. A method for using the library of claim 1 for identifying or designing enzymes or small molecule binders.
Provisional Applications (1)
Number Date Country
63300178 Jan 2022 US