Biological and Biophysical Methods for Evaluation of Inhibitors of Sortase A in Staphylococcus aureus: An Overview

• Published in: Cell biochemistry and function Vol. 42; no. 8; pp. e70002 - n/a
• Main Authors: Dewan, Debajit, Basu, Aaheli, Dolai, Debraj, Pal, Sarmistha
• Format: Journal Article
• Language: English
• Published: England Wiley Subscription Services, Inc 01.12.2024
• Subjects: Aminoacyltransferases - antagonists & inhibitors; Aminoacyltransferases - metabolism; Animals; Anti-Bacterial Agents - chemistry; Anti-Bacterial Agents - pharmacology; Antibiotic resistance; Antibiotics; Antimicrobial resistance; anti‐virulence; Assaying; Bacterial Proteins - antagonists & inhibitors; Bacterial Proteins - metabolism; biofilm; Biofilms; Biofilms - drug effects; Cell viability; Cysteine Endopeptidases - metabolism; Enzyme Inhibitors - chemistry; Enzyme Inhibitors - pharmacology; Exopolysaccharides; Fire resistance; Humans; In vivo methods and tests; Inhibitors; Laboratory tests; Peptides; Pharmacology; Sortase; sortase A; Spectroscopy; Staphylococcus aureus; Staphylococcus aureus - drug effects; Staphylococcus aureus - enzymology; Surface plasmon resonance; Therapeutic targets; Thermophoresis; Virulence; anti‐virulence; sortase A; antimicrobial resistance; Staphylococcus aureus; biofilm
• ISSN: 0263-6484; 1099-0844; 1099-0844
• DOI: 10.1002/cbf.70002

ABSTRACT Staphylococcus aureus, one of the most notorious pathogens, develops antibiotic resistance by the formation of a thick layer of exopolysaccharides known as biofilms. Sortase A, a transpeptidase responsible for biofilm formation and attachment to the host surface, has emerged as an important drug target for development of anti‐virulence agents. A number of sortase A inhibitors, both peptide and non‐peptides are reported which involved the use of several experiments which may provide insights regarding binding affinity, specificity, safety, and efficacy of ligands. In this review, we focus on the principles, pros and cons, and the type of information obtained from biophysical (FRET assay, Microscale Thermophoresis, Surface Plasmon Resonance, CD spectroscopy etc.) and biological (cell viability assay, biofilm formation assay, CLSM, western blot analysis, in vivo characterization on mice etc.) methods for estimation of probable sortase A inhibitors, which might be helpful to the researchers who might be interested to delve into the development of sortase A inhibitors as a drug, to address the burning question of antimicrobial resistance (AMR). Summary One of the most well‐known infectious agents, Staphylococcus aureus, becomes resistant to antibiotics by forming thick exopolysaccharide layers called biofilms. The transpeptidase sortase A, which is involved in the formation of biofilms and their adhesion to the host surface, has gained attention as a promising target for the development of anti‐virulence agents. Most of the commonly adopted biological and biophysical experiments for the estimation of inhibition of sortase A and biofilm formation have been discussed in one place. This work elucidating the routinely applied laboratory tests for evaluation of sortase A inhibitors, will help researchers to open up new avenues toward the development of therapeutic agents for drug‐resistant S. aureus.