Antimicrobial Synergy of a Ribonuclease and a Peptide Secreted by Human Cells

Eller, Chelcie H.; Raines, Ronald T.

doi:10.1021/acsinfecdis.0c00594

“…LL-37 and RNase 1 have been shown to act synergistically to kill E. coli . (Eller 2020) RNases have been tested in clinical trials as chemotherapeutics for the treatment of cancer. (Ardelt 2009)…”

Section: Discussionmentioning

confidence: 99%

“…LL-37 and RNase 1 have been shown to act synergistically to kill E. coli. (Eller 2020) RNases have been tested in clinical trials as chemotherapeutics for the treatment of cancer. (Ardelt 2009) Further studies are warranted to determine whether these results could be generalized to antimicrobial peptide-nuclease combinations, as might be suggested by the presence of DNA in biofilms.…”

Section: Discussionmentioning

confidence: 99%

Enhancement of the Activity of the Antimicrobial Peptides HNP1 and LL-37 by Bovine Pancreatic Ribonuclease A

Ericksen

¹

2022

Preprint

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Background: HNP1, LL-37, and HBD1 are antimicrobial against Escherichia coli ATCC 25922 at the standard inoculum but less active at higher inocula. Methods: The virtual colony count (VCC) microbiological assay was adapted for high inocula and the addition of yeast tRNA and bovine pancreatic ribonuclease A (RNase). 96-well plates were read for 12 hours in a Tecan Infinite M1000 plate reader and photographed under 10x magnification. Results: Adding tRNA 1:1 to HNP1 at the standard inoculum almost completely abrogated activity. Adding RNase 1:1 to HNP1 at the standard inoculum of 5x105 CFU/mL did not enhance activity. Increasing the inoculum to 6.25x107 CFU/mL almost abrogated HNP1 activity. However, adding RNase 25:1 to HNP1 enhanced activity. Adding both tRNA and RNase resulted in enhanced activity, indicating that the enhancement effect of RNase overwhelms the inhibiting effect of tRNA when both are present. HBD1 activity at the standard inoculum was almost completely abrogated by the addition of tRNA, but LL-37 activity was only slightly inhibited by tRNA. At the high inoculum, LL-37 activity was enhanced by RNase. HBD1 activity was not enhanced by RNase. RNase was not antimicrobial in the absence of antimicrobial peptides. Cell clumps were observed at the high inoculum in the presence of all three antimicrobial peptides and at the standard inoculum in the presence of HNP1+tRNA. Conclusions: Antimicrobial peptide-ribonuclease combinations have the potential to be active against high cell concentrations and biofilms, conditions where the antimicrobial agent alone is relatively ineffective.

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“…The combination of an antimicrobial peptide with a ribonuclease could be regarded as a novel invention that could possibly be used as a therapy to treat bacterial infections. LL-37 and RNase 1 have been shown to act synergistically to kill E. coli (Eller 2020). RNases have been tested in clinical trials as chemotherapeutics for the treatment of cancer (Ardelt 2009).…”

Section: Discussionmentioning

confidence: 99%

Enhancement of the activity of the antimicrobial peptides HNP1 and LL-37 by bovine pancreatic ribonuclease A

Ericksen

¹

2022

F1000Res

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Background: HNP1, LL-37, and HBD1 are antimicrobial against Escherichia coli ATCC 25922 at the standard inoculum but less active at higher inocula. Methods: The virtual colony count (VCC) microbiological assay was adapted for high inocula and the addition of yeast tRNA and bovine pancreatic ribonuclease A (RNase). 96-well plates were read for 12 hours in a Tecan Infinite M1000 plate reader and photographed under 10x magnification. Results: Adding tRNA 1:1 to HNP1 at the standard inoculum almost completely abrogated activity. Adding RNase 1:1 to HNP1 at the standard inoculum of 5x105 CFU/mL did not enhance activity. Increasing the inoculum to 6.25x107 CFU/mL almost abrogated HNP1 activity. However, adding RNase 25:1 to HNP1 enhanced activity. Adding both tRNA and RNase resulted in enhanced activity, indicating that the enhancement effect of RNase overwhelms the inhibiting effect of tRNA when both are present. HBD1 activity at the standard inoculum was almost completely abrogated by the addition of tRNA, but LL-37 activity was only slightly inhibited by tRNA. At the high inoculum, LL-37 activity was enhanced by RNase. HBD1 activity was not enhanced by RNase. RNase was not antimicrobial in the absence of antimicrobial peptides. Cell clumps were observed at the high inoculum in the presence of all three antimicrobial peptides and at the standard inoculum in the presence of HNP1+tRNA. Conclusions: Antimicrobial peptide-ribonuclease combinations have the potential to be active against high cell concentrations and biofilms, conditions where the antimicrobial agent alone is relatively ineffective.

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“…RNase 1 (UniProtKB P07998), which is the human homologue of RNase A, has demonstrable utility in biomedical contexts. In particular, an engineered variant of RNase 1 exhibits specific toxicity for cancer cells and is in clinical trials as a chemotherapeutic agent. , RNase 1 also has considerable antiviral activity and manifests antibacterial activity in the presence of a pore-forming peptide . To date, however, RNase S has not been employed in the clinic.…”

mentioning

confidence: 99%

“…16,17 RNase 1 also has considerable antiviral activity 18 and manifests antibacterial activity in the presence of a pore-forming peptide. 19 To date, however, RNase S has not been employed in the clinic. Indeed, such applications are discouraged by the potential immunogenicity of the bovine-derived complex in humans.…”

mentioning

confidence: 99%

Semisynthesis of Human Ribonuclease–S

Sayers

¹

,

Wralstad

²

,

Raines

³

2020

Bioconjugate Chem.

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Since its conception, the ribonuclease S complex (RNase S) has led to historic discoveries in protein chemistry, enzymology, and related fields. Derived by the proteolytic cleavage of a single peptide bond in bovine pancreatic ribonuclease (RNase A), RNase S serves as a convenient and reliable model system for incorporating unlimited functionality into an enzyme. Applications of the RNase S system in biomedicine and biotechnology have, however, been hindered by two shortcomings: (1) the bovine-derived enzyme could elicit an immune response in humans, and (2) the complex is susceptible to dissociation. Here, we have addressed both limitations in the first semisynthesis of an RNase S conjugate derived from human pancreatic ribonuclease and stabilized by a covalent interfragment cross-link. We anticipate that this strategy will enable unprecedented applications of the "RNase-S" system.

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Antimicrobial Synergy of a Ribonuclease and a Peptide Secreted by Human Cells

Cited by 12 publications

References 53 publications

Enhancement of the Activity of the Antimicrobial Peptides HNP1 and LL-37 by Bovine Pancreatic Ribonuclease A

Enhancement of the Activity of the Antimicrobial Peptides HNP1 and LL-37 by Bovine Pancreatic Ribonuclease A

Enhancement of the activity of the antimicrobial peptides HNP1 and LL-37 by bovine pancreatic ribonuclease A

Semisynthesis of Human Ribonuclease–S

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