An E. coli display method for characterization of peptide–sensor kinase interactions

Brink, Kathryn R.; Hunt, Maxwell; Mu, Andrew; Groszman, Ken; Hoang, Ky V.; Lorch, Kevin P; Pogostin, Brett H; Tabor, Jeffrey J.

doi:10.1038/s41589-022-01207-z

Cited by 8 publications

(5 citation statements)

References 59 publications

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“…Interestingly, the magnesium replacement mechanism cannot explain PhoQ activation by antimicrobial peptides in some cases. For example, a recent study revealed that antimicrobial peptides with a β-sheet structure or a neutral charge could also activate PhoQ ( 39 ). These atypical PhoQ activators do not possess the properties to interact with the PhoQ acidic cluster.…”

Section: Discussionmentioning

confidence: 99%

The inhibitory mechanism of a small protein reveals its role in antimicrobial peptide sensing

Jiang,

Steup,

Kippnich

et al. 2023

Proc. Natl. Acad. Sci. U.S.A.

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A large number of small membrane proteins have been uncovered in bacteria, but their mechanism of action has remained mostly elusive. Here, we investigate the mechanism of a physiologically important small protein, MgrB, which represses the activity of the sensor kinase PhoQ and is widely distributed among enterobacteria. The PhoQ/PhoP two-component system is a master regulator of the bacterial virulence program and interacts with MgrB to modulate bacterial virulence, fitness, and drug resistance. A combination of cross-linking approaches with functional assays and protein dynamic simulations revealed structural rearrangements due to interactions between MgrB and PhoQ near the membrane/periplasm interface and along the transmembrane helices. These interactions induce the movement of the PhoQ catalytic domain and the repression of its activity. Without MgrB, PhoQ appears to be much less sensitive to antimicrobial peptides, including the commonly used C18G. In the presence of MgrB, C18G promotes MgrB to dissociate from PhoQ, thus activating PhoQ via derepression. Our findings reveal the inhibitory mechanism of the small protein MgrB and uncover its importance in antimicrobial peptide sensing.

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Section: Discussionmentioning

confidence: 99%

The inhibitory mechanism of a small protein reveals its role in antimicrobial peptide sensing

Jiang,

Steup,

Kippnich

et al. 2023

Proc. Natl. Acad. Sci. U.S.A.

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“…Both hypotheses, however, are not in conflict with each other but rather describe two aspects of PhoQ sensing CAMPs under different magnesium conditions. Furthermore, a recent study revealed that antimicrobial peptides with ß-sheet structure or neutral charge could also activate PhoQ [54]. These newly found PhoQ activators do not possess the properties to interact with the PhoQ acidic patch.…”

Section: Discussionmentioning

confidence: 99%

The inhibitory mechanism of a small protein reveals its role in antimicrobial peptide sensing

Jiang

Steup

Kippnich

et al. 2022

Preprint

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A large number of small membrane proteins have been discovered in bacteria, but their mechanism of action has remained mostly elusive. Here, we investigate the mechanism of a physiologically important small protein, MgrB, which represses the activity of the sensor kinase PhoQ and is widely distributed among enterobacteria. The PhoQ/PhoP two-component system is a master regulator of the bacterial virulence program and interacts with MgrB to modulate bacterial virulence, fitness, and drug resistance. A combination of crosslinking approaches with functional assays and protein dynamic simulations revealed structural rearrangements due to interactions between MgrB and PhoQ near the membrane/periplasm interface and along the transmembrane helices. These interactions induce the movement of the PhoQ catalytic domain and the repression of its activity. In the absence of MgrB, PhoQ appears to be insensitive to antimicrobial peptides, including the commonly used C18G. MgrB mediates the sensing of C18G by dissociating from and thus derepressing PhoQ. Our findings reveal the inhibitory mechanism of the small protein MgrB and uncover its importance in antimicrobial peptide sensing.

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“…The protease could directly or indirectly activate regulated intramembrane proteolysis [50]. Or, it could cleave a pro-peptide that activates a two-component regulatory system [51]. The benefit of either of those approaches is that an exogenous small molecule would not be required for the continued survival and replication of the vector.…”

Section: Other Organs and Organ Systemsmentioning

confidence: 99%

Encapsulated Bacteria with a Light-Repressed Deadman Switch for Liver Gene Delivery

Renteln

2024

Preprint

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Adeno-associated viruses (AAVs) have been used for liver gene therapy. Hemgenix and Roctavian are AAV-based treatments for hemophilia B and A, respectively. They cost $3 million and $2.8 million per dose, respectively. While AAV vectors may eventually be cheaper to mass produce, a bacteria-based DNA delivery system might be much cheaper for patients. Also, “bactofection” would allow for the delivery of much larger DNA packages. Such a bacterial system may now be possible, and a prototype for the liver could possibly be developed immediately.

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An E. coli display method for characterization of peptide–sensor kinase interactions

Cited by 8 publications

References 59 publications

The inhibitory mechanism of a small protein reveals its role in antimicrobial peptide sensing

The inhibitory mechanism of a small protein reveals its role in antimicrobial peptide sensing

The inhibitory mechanism of a small protein reveals its role in antimicrobial peptide sensing

Encapsulated Bacteria with a Light-Repressed Deadman Switch for Liver Gene Delivery

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