Chimeric antibacterial polypeptides
Provided herein are antibacterial compositions and methods of making and using the compositions.
1. A chimeric polypeptide comprising:
(i) a muralytic domain (MD) from a virion-associated muralytic enzyme or a variant thereof capable of lysing chloroform-treated Pseudomonas aeruginosa bacteria; and
(ii) a membrane traversing domain (MTD) having a DAS profile between 1.5 and 3, and capable of traversing the outer membrane of Pseudomonas aeruginosa bacteria.
2. The chimeric polypeptide of claim 1 , wherein 1-100 nmol of the MD in the absence of the MTD lyses at least 50% of 10 7 chloroform-treated Pseudomonas aeruginosa bacteria in a CFU drop assay.
3. A chimeric polypeptide of claim 1 , wherein:
(i) the MD comprises a sequence of amino acids 737-875 of SEQ ID NO:2, or a variant thereof capable of lysing chloroform-treated gram negative bacteria; and
(ii) the MTD comprises amino acids 16-39 of SEQ ID NO:4, or a variant thereof with a DAS profile between 1.2 and 2.6.
4. The chimeric polypeptide of claim 1 , wherein 1-100 nmol of the chimeric polypeptide lyses at least 50% of 10 7 Gram negative bacteria in a CFU drop assay.
5. The chimeric polypeptide of claim 1 , wherein said MD comprises a sequence of amino acids 737-875 of SEQ ID NO:2, or a variant with 1, 2, or 3 of the methionine amino acids substituted with non-methionine amino acids.
6. The chimeric polypeptide of claim 1 , wherein said MD comprises a sequence of amino acids 683-889 of SEQ ID NO:2, or a variant with 1-7 of the methionine amino acids substituted with non-methionine amino acids.
7. The chimeric polypeptide of claim 1 , wherein the MTD comprises a sequence of amino acids 16-39 of SEQ ID NO:4, or a variant with 1-6 hydrophobic amino acids substituted with amino acids with a hydropathy score of −2 or lower.
8. The chimeric polypeptide of claim 7 , wherein V20, V22, and 124 of SEQ ID NO:4 are substituted with amino acids with a hydropathy score of −2 or lower.
9. The chimeric polypeptide of claim 1 , wherein the DAS profile of the MTD is under 2.5.
10. The chimeric polypeptide of claim 1 , wherein the DAS profile of the chimeric polypeptide is under 2.5.
11. The chimeric polypeptide of claim 1 , wherein the MTD and MD are joined by a linker comprising 3-5 positively charged amino acids.
12. The chimeric polypeptide of claim 1 , wherein the MD comprises a sequence at least 90% identical to amino acids 737-875 of SEQ ID NO:2 and the MTD comprises a sequence at least 80% identical to amino acids 16-39 of SEQ ID NO:4.
13. The chimeric polypeptide of claim 1 , wherein the chimeric polypeptide is attached to polyethylene glycol.
14. A pharmaceutical composition comprising the chimeric polypeptide of claim 1 and a pharmaceutically acceptable excipient.
15. A method of inhibiting bacterial cell growth in an individual comprising administering the pharmaceutical composition of claim 14 to the individual.
16. The method of claim 15 , wherein the bacteria are selected from the group consisting of Pseudomonas aeruginosa, Klebsiella pneumoniae, Escherichia coli, Klebsiella pneumoniae, Acinetobacter baumanii, Salmonella typhimurium, Salmonella infantis, Shigella, Proteus mirabilis , and Burkholderia thailandensis.
17. An antibacterial formulation comprising the chimeric polypeptide of claim 15 and an agent that reduces oxidation.
18. A method of inhibiting bacterial cell growth in an environment comprising applying the chimeric polypeptide of claim 1 to the environment.
19. The method of claim 18 , wherein the bacteria are selected from the group consisting of Pseudomonas aeruginosa, Klebsiella pneumoniae, Escherichia coli, Klebsiella pneumoniae, Acinetobacter baumanii, Salmonella typhimurium, Salmonella infantis, Shigella, Proteus mirabilis , and Burkholderia thailandensis.