The importance of the PapR7 C-terminus and amide protons in mediating quorum sensing in Bacillus cereus

Michael Gorgan; Shahar Vanunu Ofri; Emilee R. Engler; Avishag Yehuda; Elizabeth Hutnick; Zvi Hayouka; Michael A. Bertucci

doi:10.1016/j.resmic.2023.104139

The importance of the PapR₇ C-terminus and amide protons in mediating quorum sensing in Bacillus cereus

Michael Gorgan, Shahar Vanunu Ofri, Emilee R. Engler, Avishag Yehuda, Elizabeth Hutnick, Zvi Hayouka^*, Michael A. Bertucci^*

^*Corresponding author for this work

Institute of Biochemistry, Food Science and Nutrition

Research output: Contribution to journal › Article › peer-review

Abstract

The opportunistic human pathogen Bacillus cereus controls the expression of key infection-promoting phenotypes using bacterial quorum sensing (QS). QS signal transduction within the species is controlled by an autoinducing peptide, PapR₇, and its cognate receptor, PlcR, indicating that the PlcR:PapR interface is a prime target for QS inhibitor development. The C-terminal region of the peptide (PapR₇; ADLPFEF) has been successfully employed as a scaffold to develop potent QS modulators. Despite the noted importance of the C-terminal carboxylate and amide protons in crystallographic data, their role in QS activity has yet to be explored. In this study, an N-methyl scan of PapR₇ was conducted in conjunction with a C-terminal modification of previously identified B. cereus QS inhibitors. The results indicate that the amide proton at Glu6 and the C-terminal carboxylate are important for effective QS inhibition of the PlcR regulon. Through β-galactosidase and hemolysis assays, a series of QS inhibitors were discovered, including several capable of inhibiting QS with nanomolar potency. These inhibitors, along with the structure–activity data reported, will serve as valuable tools for disrupting the B. cereus QS pathway towards developing novel anti-infective strategies.

Original language	American English
Article number	104139
Journal	Research in Microbiology
DOIs	https://doi.org/10.1016/j.resmic.2023.104139
State	Accepted/In press - 2023

Bibliographical note

Funding Information:
We would like to thank the Lafayette College and Moravian University chemistry departments for financial support. S.H.O would like to thank the Robert H. Smith Faculty of Agriculture, Food and Environment of the Hebrew University of Jerusalem for her MSc fellowship.

Funding Information:
We would like to thank the Lafayette College and Moravian University chemistry departments for financial support. S.H.O would like to thank the Robert H. Smith Faculty of Agriculture, Food and Environment of the Hebrew University of Jerusalem for her MSc fellowship.

Publisher Copyright:
© 2023 Institut Pasteur

Keywords

Bacillus cereus group
Peptide structure–activity relationships
PlcR antagonists
Quorum sensing

Access to Document

10.1016/j.resmic.2023.104139

Cite this

@article{ac01614c3cea4d4db9f0d8690ab8bf25,

title = "The importance of the PapR7 C-terminus and amide protons in mediating quorum sensing in Bacillus cereus",

abstract = "The opportunistic human pathogen Bacillus cereus controls the expression of key infection-promoting phenotypes using bacterial quorum sensing (QS). QS signal transduction within the species is controlled by an autoinducing peptide, PapR7, and its cognate receptor, PlcR, indicating that the PlcR:PapR interface is a prime target for QS inhibitor development. The C-terminal region of the peptide (PapR7; ADLPFEF) has been successfully employed as a scaffold to develop potent QS modulators. Despite the noted importance of the C-terminal carboxylate and amide protons in crystallographic data, their role in QS activity has yet to be explored. In this study, an N-methyl scan of PapR7 was conducted in conjunction with a C-terminal modification of previously identified B. cereus QS inhibitors. The results indicate that the amide proton at Glu6 and the C-terminal carboxylate are important for effective QS inhibition of the PlcR regulon. Through β-galactosidase and hemolysis assays, a series of QS inhibitors were discovered, including several capable of inhibiting QS with nanomolar potency. These inhibitors, along with the structure–activity data reported, will serve as valuable tools for disrupting the B. cereus QS pathway towards developing novel anti-infective strategies.",

keywords = "Bacillus cereus group, Peptide structure–activity relationships, PlcR antagonists, Quorum sensing",

author = "Michael Gorgan and {Vanunu Ofri}, Shahar and Engler, {Emilee R.} and Avishag Yehuda and Elizabeth Hutnick and Zvi Hayouka and Bertucci, {Michael A.}",

note = "Funding Information: We would like to thank the Lafayette College and Moravian University chemistry departments for financial support. S.H.O would like to thank the Robert H. Smith Faculty of Agriculture, Food and Environment of the Hebrew University of Jerusalem for her MSc fellowship. Funding Information: We would like to thank the Lafayette College and Moravian University chemistry departments for financial support. S.H.O would like to thank the Robert H. Smith Faculty of Agriculture, Food and Environment of the Hebrew University of Jerusalem for her MSc fellowship. Publisher Copyright: {\textcopyright} 2023 Institut Pasteur",

year = "2023",

doi = "10.1016/j.resmic.2023.104139",

language = "American English",

journal = "Research in Microbiology",

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AU - Gorgan, Michael

AU - Vanunu Ofri, Shahar

AU - Engler, Emilee R.

AU - Yehuda, Avishag

AU - Hutnick, Elizabeth

AU - Hayouka, Zvi

AU - Bertucci, Michael A.

N1 - Funding Information: We would like to thank the Lafayette College and Moravian University chemistry departments for financial support. S.H.O would like to thank the Robert H. Smith Faculty of Agriculture, Food and Environment of the Hebrew University of Jerusalem for her MSc fellowship. Funding Information: We would like to thank the Lafayette College and Moravian University chemistry departments for financial support. S.H.O would like to thank the Robert H. Smith Faculty of Agriculture, Food and Environment of the Hebrew University of Jerusalem for her MSc fellowship. Publisher Copyright: © 2023 Institut Pasteur

PY - 2023

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N2 - The opportunistic human pathogen Bacillus cereus controls the expression of key infection-promoting phenotypes using bacterial quorum sensing (QS). QS signal transduction within the species is controlled by an autoinducing peptide, PapR7, and its cognate receptor, PlcR, indicating that the PlcR:PapR interface is a prime target for QS inhibitor development. The C-terminal region of the peptide (PapR7; ADLPFEF) has been successfully employed as a scaffold to develop potent QS modulators. Despite the noted importance of the C-terminal carboxylate and amide protons in crystallographic data, their role in QS activity has yet to be explored. In this study, an N-methyl scan of PapR7 was conducted in conjunction with a C-terminal modification of previously identified B. cereus QS inhibitors. The results indicate that the amide proton at Glu6 and the C-terminal carboxylate are important for effective QS inhibition of the PlcR regulon. Through β-galactosidase and hemolysis assays, a series of QS inhibitors were discovered, including several capable of inhibiting QS with nanomolar potency. These inhibitors, along with the structure–activity data reported, will serve as valuable tools for disrupting the B. cereus QS pathway towards developing novel anti-infective strategies.

AB - The opportunistic human pathogen Bacillus cereus controls the expression of key infection-promoting phenotypes using bacterial quorum sensing (QS). QS signal transduction within the species is controlled by an autoinducing peptide, PapR7, and its cognate receptor, PlcR, indicating that the PlcR:PapR interface is a prime target for QS inhibitor development. The C-terminal region of the peptide (PapR7; ADLPFEF) has been successfully employed as a scaffold to develop potent QS modulators. Despite the noted importance of the C-terminal carboxylate and amide protons in crystallographic data, their role in QS activity has yet to be explored. In this study, an N-methyl scan of PapR7 was conducted in conjunction with a C-terminal modification of previously identified B. cereus QS inhibitors. The results indicate that the amide proton at Glu6 and the C-terminal carboxylate are important for effective QS inhibition of the PlcR regulon. Through β-galactosidase and hemolysis assays, a series of QS inhibitors were discovered, including several capable of inhibiting QS with nanomolar potency. These inhibitors, along with the structure–activity data reported, will serve as valuable tools for disrupting the B. cereus QS pathway towards developing novel anti-infective strategies.

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