(US20190040107) Astexin peptides 机翻标题: 暂无翻译,请尝试点击翻译按钮。

源语言标题
(US20190040107) Astexin peptides
公开号/公开日
US20190040107 / 2019-02-07
申请号/申请日
US16/057,292 / 2018-08-07
发明人
LINK A JAMESMAKSIMOV MIKHAIL O;
申请人
PRINCETON UNIVERSITY;
主分类号
IPC分类号
A01N-063/02 A62D-003/33 A62D-101/43 C02F-001/28 C02F-101/20 C07K-014/195
摘要
(US20190040107) Astexin-1 is highly polar, in contrast to many lasso peptides that are primarily hydrophobic, and has modest antimicrobial activity against Caulobacter crescentus, a bacterium related to Asticcacaulis excentricus. The solution structure of astexin-1 was determined, revealing a unique topology that is stabilized by hydrogen bonding between segments of the peptide. Astexins-2 and -3 are intracellular lasso peptides.
机翻摘要
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地址
代理人
代理机构
;
优先权号
2012US-61695772 2013US-61839493 2013WO-US57203 2015US-14424617 2018US-16057292
主权利要求
(US20190040107) What is claimed is: 1. A substantially purified Astexin-1 peptide comprising the amino acid sequence   H2N-X1-X2-X3-X4-X5-X6-X7-X8-X9-X10-X11-X12-X13-X14-X15-X16-X17-X18-X19-X20-X21-X22-X23-X24-COOH, wherein    X1 is absent or a peptide sequence or a naturally or non-naturally occurring amino acid;    X2 is absent or G or a small naturally or non-naturally occurring amino acid;    X3 is absent or L or a hydrophobic or a small naturally or non-naturally occurring amino acid;    X4 is absent or S or a polar naturally or non-naturally occurring amino acid;    X5 is absent or Q or a polar or acidic naturally or non-naturally occurring amino acid;    X6 is absent or G or a small naturally or non-naturally occurring amino acid;    X7 is absent or V or a hydrophobic or a small naturally or non-naturally occurring amino acid;    X8 is absent or E or a hydrophobic or a small naturally or non-naturally occurring amino acid;    X9 is absent or P or a basic or a polar naturally or non-naturally occurring amino acid;    X10 is absent or D or an acidic or polar naturally or non-naturally occurring amino acid;    X11 is absent or I or a hydrophobic naturally or non-naturally occurring amino acid;    X12 is absent or G or a small naturally or non-naturally occurring amino acid;    X13 is absent or Q or a polar or basic naturally or non-naturally occurring amino acid;    X14 is absent or Tor a polar naturally or non-naturally occurring amino acid;    X15 is absent or Y or a polar or acidic naturally or non-naturally occurring amino acid;    X16 is absent or F or hydrophobic or an aromatic naturally or non-naturally occurring amino acid;    X17 is absent or E or a basic or a polar naturally or non-naturally occurring amino acid;    X18 is absent or E or absent or a polar or acidic naturally or non-naturally occurring amino acid;    X19 is absent or S or a polar naturally or non-naturally occurring amino acid;    X20 is absent or R or a basic or polar naturally or non-naturally occurring amino acid;    X21 is absent or I or a hydrophobic naturally or non-naturally occurring amino acid;    X22 is absent or N or a polar or basic naturally or non-naturally occurring amino acid;    X23 is absent or Q or a polar or basic naturally or non-naturally occurring amino acid;    X24 is absent or D or a hydrophobic or a small naturally or non-naturally occurring amino acid; and    X25 is absent or a naturally or non-naturally occurring amino acid or a small peptide,   or a pharmaceutically acceptable salt thereof. 2. A substantially purified Astexin-2 peptide comprising the amino acid sequence   H2N-Y1-Y2-Y3-Y4-Y5-Y6-Y7-Y8-Y9-Y10-Y11-Y12-Y13-Y14-Y15-Y16-Y17-Y18-Y19-Y20-Y21-Y22-Y23-Y24-COOH, wherein    Y1 is absent or a peptide sequence or a naturally or non-naturally occurring amino acid;    Y2 is absent or G or a small naturally or non-naturally occurring amino acid;    Y3 is absent or L or a hydrophobic or a small naturally or non-naturally occurring amino acid;    Y4 is absent or T or a polar naturally or non-naturally occurring amino acid;    Y5 is absent or Q or a polar or acidic naturally or non-naturally occurring amino acid;    Y6 is absent or I or a hydrophobic naturally or non-naturally occurring amino acid;    Y7 is absent or Q or a polar or acidic naturally or non-naturally occurring amino acid;    Y8 is absent or A or a hydrophobic or a small naturally or non-naturally occurring amino acid;    Y9 is absent or L or a hydrophobic or a small naturally or non-naturally occurring amino acid;    Y10 is absent or D or a hydrophobic or a small or an acidic naturally or non-naturally occurring amino acid;    Y11 is absent or S or a polar naturally or non-naturally occurring amino acid;    Y12 is absent or V or a hydrophobic naturally or non-naturally occurring amino acid;    Y13 is absent or S or a polar naturally or non-naturally occurring amino acid;    Y14 is absent or G or a small naturally or non-naturally occurring amino acid;    Y15 is absent or Q or a polar or acidic naturally or non-naturally occurring amino acid;    Y16 is absent or F or hydrophobic or an aromatic naturally or non-naturally occurring amino acid;    Y17 is absent or R or a basic or a polar naturally or non-naturally occurring amino acid;    Y18 is absent or D or absent or a polar or an acidic or a polar naturally or non-naturally occurring amino acid;    Y19 is absent or Q or basic or a polar naturally or non-naturally occurring amino acid;    Y20 is absent or L or a hydrophobic or a small naturally or non-naturally occurring amino acid;    Y21 is absent or G or a small naturally or non-naturally occurring amino acid;    Y22 is absent or L or a hydrophobic or a small naturally or non-naturally occurring amino acid;    Y23 is absent or S or a polar naturally or non-naturally occurring amino acid    Y24 is absent or A or a hydrophobic or a small naturally or non-naturally occurring amino acid;    Y25 is absent or D or a polar or an acidic or a polar naturally or non-naturally occurring amino acid,   or a pharmaceutically acceptable salt thereof. 3. A substantially purified Astexin-3 peptide comprising the amino acid sequence   H2N-Z1-Z2-Z3-Z4-Z5-Z6-Z7-Z8-Z9-Z10-Z11-Z12-Z13-Z14-Z15-Z16-Z17-Z18-Z19-Z20- Z21-Z22-Z23-Z24-Z25-Z26-COOH,   wherein    Z1 is absent or a peptide sequence or a naturally or non-naturally occurring amino acid;    Z2 is absent or G or a small naturally or non-naturally occurring amino acid;    Z3 is absent or P or a naturally or non-naturally occurring amino acid;    Z4 is absent or T or a polar naturally or non-naturally occurring amino acid;    Z5 is absent or P or a naturally or non-naturally occurring amino acid;    Z6 is absent or M or a hydrophobic naturally or non-naturally occurring amino acid;    Z7 is absent or V or a hydrophobic naturally or non-naturally occurring amino acid;    Z8 is absent or G or a small naturally or non-naturally occurring amino acid;    Z9 is absent or L or a hydrophobic naturally or non-naturally occurring amino acid;    Z10 is absent or D or a small or an acidic naturally or non-naturally occurring amino acid;    Z11 is absent of S or a small or polar naturally or non-naturally occurring amino acid;    Z12 is absent or V or a hydrophobic naturally or non-naturally occurring amino acid;    Z13 is absent or S or a small or polar naturally or non-naturally occurring amino acid;    Z14 is absent or G or a small naturally or non-naturally occurring amino acid;    Z15 is absent or Q or a basic or a polar naturally or non-naturally occurring amino acid;    Z16 is absent or Y or a hydrophobic or an aromatic naturally or non-naturally occurring amino acid;    Z17 is absent or W or a hydrophobic or an aromatic naturally or non-naturally occurring amino acid;    Z18 is absent or D or a small or an acidic naturally or non-naturally occurring amino acid;    Z19 is absent or Q or a basic or a polar naturally or non-naturally occurring amino acid;    Z20 is absent or H or a basic or polar naturally or non-naturally occurring amino acid;    Z21 is absent or A or a small hydrophobic naturally or non-naturally occurring amino acid;    Z22 is absent or P or a naturally or non-naturally occurring amino acid;    Z23 is absent or L or a hydrophobic naturally or non-naturally occurring amino acid;    Z24 is absent or A or a small or hydrophobic naturally or non-naturally occurring amino acid;    Z25 is absent or D or a polar or acidic naturally or non-naturally occurring amino acid; and    Z26 is absent or a peptide or a polar or acidic naturally or non-naturally occurring amino acid. 4. The peptide of claim 1, 2, or 3, wherein the peptide is a lasooed peptide. 5. The peptide of claim 4, wherein the G at X2 is covalently bound to D at X10. 6. The peptide of claim 1, wherein the peptide comprises the sequence GLSQGVEPDIGQTYFEESRINQD (SEQ ID NO:3). 7. The peptide of claim 1, wherein X1 is MHTPIISTTVQPKT (SEQ ID NO:4). 8. The peptide of claim 1, 2, or 3, wherein the peptide is less than 50 amino acids. 9. The peptide of claim 1, 2, or 3, wherein the peptide is less than 35 amino acids. 10. The peptide of claim 1, 2, or 3, wherein the peptide is a substrate for an isopeptidase. 11. The peptide of claim 10, wherein the peptide is a substrate for an AtxE2 isopeptidase. 12. The peptide of claim 1, 2, or 3, wherein the peptide is isolated from a cell that contains a naturally occurring astexin peptide. 13. The peptide of claim 1, 2, or 3, wherein the peptide is isolated from an astexin peptide recombinantly produced in a cell. 14. The peptide of claim 13, wherein the cell is a prokaryotic cell. 15. The peptide of claim 13, wherein the cell is a eukaryotic cell. 16. The peptide of claim 1, 2, or 3, wherein the peptide is chemically synthesized in vitro. 17. A non-naturally occurring polynucleotide sequence encoding the peptide of claim 1, 2, or 3. 18. A vector comprising the polynucleotide of claim 17. 19. A cell containing the vector of claim 18. 20. The cell of claim 19, wherein the cell is a prokaryotic cell. 21. The cell of claim 18, wherein the cell is an E. coli cell. 22. The cell of claim 19, wherein the cell is an Asticcacaulis excentricus cell. 23. An astexin library comprising a plurality of vectors encoding an astexin peptide, wherein the vectors include an insertion of one to five codons of a polynucleotide sequence encoding a non-astexin polypeptide. 24. The library of claim 23, wherein the insertion is in a loop or ring of the astexin peptide. 25. The library of claim 23, wherein the insertion is 3-4 codons of a non-astexin polypeptide-encoding region of the polynucleotide. 26. A method of expressing a peptide, the method comprising culturing a cell expressing a polynucleotide encoding an astexin peptide under conditions allowing for expression of the astexin peptide. 27. The method of claim 26, further comprising recovering the astexin peptide. 28. A method of removing a toxic substance from a sample suspected of containing the toxic substance, the method comprising   contacting the sample with an astexin peptide under conditions that allow for formation of a complex between the toxic substance and the peptide, and   removing the complex from the sample,   thereby removing the toxic substance from the sample. 29. The method of claim 28, wherein the sample is a soil sample. 30. The method of claim 28, wherein the sample is a water sample. 31. The method of claim 28, wherein the toxic substance is a metal. 32. The method of claim 31, wherein the metal is cadmium or lead. 33. A method of inhibiting the growth of a microbe, the method comprising contacting the microbe with an astexin peptide in an amount sufficient to inhibit growth of the microbe. 34. The method of claim 33, wherein the astexin peptide binds heavy metals in an amount sufficient to inhibit growth of the microbe. 35. The method of claim 33, wherein the microbe is a prokaryote. 36. The method of claim 33, wherein the microbe is a eukaryote. 37. A method of purifying a protein, the method comprising   heating a solution containing an astexin fusion protein, the fusion protein comprising an astexin peptide and a second protein moiety,   centrifuging the heated solution,   recovering the astexin fusion protein;   cleaving the fusion protein with a lasso protease to separate the astexin peptide and the second protein moiety, and   isolating the second protein moiety,   thereby purifying the protein. 38. A substantially purified AtxE2 polypeptide at least 85% identical to the ATEX2 polypeptide sequence of SEQ ID NO: 1. 39. The polypeptide of claim 38, wherein the polypeptide includes a catalytic serine residue at position 527 when numbered with respect to the corresponding ATEX2 polypeptide sequence. 40. The polypeptide of claim 38, wherein the polypeptide is at least 90% identical to the ATXE2 polypeptide sequence of SEQ ID NO: 1. 41. The polypeptide of claim 38, wherein the polypeptide is 95% to 98% identical to the ATEX2 polypeptide sequence of SEQ ID NO: 1. 42. The polypeptide of claim 36, wherein the polypeptide is at least 98% identical to the ATEX2 polypeptide sequence of SEQ ID NO: 1. 43. A non-naturally occurring polynucleotide sequence encoding the polypeptide of claim 38. 44. A method of regulating expression or activity of an astexin-2 peptide, the method comprising   providing a cell containing (1) a polynucleotide encoding an astexin peptide and (2) a polynucleotide encoding an AtxE2 polypeptide;   culturing the cell under conditions allowing for expression of the astexin peptide and second moiety, and   inducing expression of an AtxE2 polypeptide under conditions that cause hydrolysis of the astexin peptide, thereby regulating expression or activity of the astexin peptide. 45. The method of claim 44, wherein the astexin-encoding polynucleotide is operably linked to a second moiety. 46. The method of claim 45, wherein the second moiety is a polypeptide or a label. 47. The method of claim 45, wherein the cell does not naturally express an Astexin-2 peptide or an AtxE2 peptide. 48. The method of claim 42, wherein the astexin-2 encoding polynucleotide and AtxE2 encoding polynucleotide are covalently linked. 49. The method of claim 42, wherein the astexin-2 encoding polynucleotide and AtxE2-encoding polynucleotide are on separate polynucleotides.
法律状态
PENDING
专利类型码
A1
国别省市代码
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