SaccuFlow: A High-Throughput Analysis Platform to Investigate Bacterial Cell Wall Interactions

Overview

Journal ACS Infect Dis

Specialties Infectious Diseases
Microbiology
Pharmacology

Date 2021 Jul 22

PMID 34291914

Citations 10

Authors

Alexis J Apostolos

Noel J Ferraro

Brianna E Dalesandro

Marcos M Pires

Affiliations

Soon will be listed here.

Abstract

Bacterial cell walls are formidable barriers that protect bacterial cells against external insults and oppose internal turgor pressure. While cell wall composition is variable across species, peptidoglycan is the principal component of all cell walls. Peptidoglycan is a mesh-like scaffold composed of cross-linked strands that can be heavily decorated with anchored proteins. The biosynthesis and remodeling of peptidoglycan must be tightly regulated by cells because disruption to this biomacromolecule is lethal. This essentiality is exploited by the human innate immune system in resisting colonization and by a number of clinically relevant antibiotics that target peptidoglycan biosynthesis. Evaluation of molecules or proteins that interact with peptidoglycan can be a complicated and, typically, qualitative effort. We have developed a novel assay platform (SaccuFlow) that preserves the native structure of bacterial peptidoglycan and is compatible with high-throughput flow cytometry analysis. We show that the assay is facile and versatile as demonstrated by its compatibility with sacculi from Gram-positive bacteria, Gram-negative bacteria, and mycobacteria. Finally, we highlight the utility of this assay to assess the activity of sortase A from against potential antivirulence agents.

Citing Articles

Identification of a family of peptidoglycan transpeptidases reveals that requires noncanonical cross-links for viability.

Bollinger K, Muh U, Ocius K, Apostolos A, Pires M, Helm R Proc Natl Acad Sci U S A. 2024; 121(34):e2408540121.

PMID: 39150786 PMC: 11348318. DOI: 10.1073/pnas.2408540121.

Noninvasive Analysis of Peptidoglycan from Living Animals.

Ocius K, Kolli S, Ahmad S, Dressler J, Chordia M, Jutras B Bioconjug Chem. 2024; 35(4):489-498.

PMID: 38591251 PMC: 11036361. DOI: 10.1021/acs.bioconjchem.4c00007.

Measurement of Accumulation of Antibiotics to Staphylococcus aureus in Phagosomes of Live Macrophages.

Kelly J, Dalesandro B, Liu Z, Chordia M, Ongwae G, Pires M Angew Chem Int Ed Engl. 2023; 63(3):e202313870.

PMID: 38051128 PMC: 10799677. DOI: 10.1002/anie.202313870.

Shedding Light on Bacterial Physiology with Click Chemistry.

Zheng Q, Chang P Isr J Chem. 2023; 63(1-2).

PMID: 37841997 PMC: 10569449. DOI: 10.1002/ijch.202200064.

High-Throughput Screening of Natural Product and Synthetic Molecule Libraries for Antibacterial Drug Discovery.

Ayon N Metabolites. 2023; 13(5).

PMID: 37233666 PMC: 10220967. DOI: 10.3390/metabo13050625.

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