scholarly journals High-Throughput Screen for Cell Wall Synthesis Network Module in Mycobacterium tuberculosis Based on Integrated Bioinformatics Strategy

2020 ◽  
Vol 8 ◽  
Author(s):  
Xizi Luo ◽  
Jiahui Pan ◽  
Qingyu Meng ◽  
Juanjuan Huang ◽  
Wenfang Wang ◽  
...  
Keyword(s):  
Cell Wall ◽  
Mycobacterium Tuberculosis ◽  
High Throughput ◽  
Network Module ◽  
High Throughput Screen ◽  
Cell Wall Synthesis

scholarly journals Drug Targeting Mycobacterium tuberculosis Cell Wall Synthesis: Development of a Microtiter Plate-Based Screen for UDP-Galactopyranose Mutase and Identification of an Inhibitor from a Uridine-Based Library

2003 ◽  
Vol 47 (1) ◽  
pp. 378-382 ◽  
Author(s):  
Michael S. Scherman ◽  
Katharine A. Winans ◽  
Richard J. Stern ◽  
Victoria Jones ◽  
Carolyn R. Bertozzi ◽  
...  
Keyword(s):  
Cell Wall ◽  
Mycobacterium Tuberculosis ◽  
Drug Targeting ◽  
Enzyme Inhibitor ◽  
Microtiter Plate ◽  
Biosynthetic Enzyme ◽  
Cell Wall Synthesis ◽  
Chemical Library ◽  
Microtiter Plate Assay ◽  
Plate Assay

ABSTRACT A microtiter plate assay for UDP-galactopyranose mutase, an essential cell wall biosynthetic enzyme of Mycobacterium tuberculosis, was developed. The assay is based on the release of tritiated formaldehyde from UDP-galactofuranose but not UDP-galactopyranose by periodate and was used to identify a uridine-based enzyme inhibitor from a chemical library.

scholarly journals A High Throughput Screen for Inhibitors of Fungal Cell Wall Synthesis

2002 ◽  
Vol 7 (4) ◽  
pp. 359-366 ◽  
Author(s):  
Jonathan M. Evans ◽  
Phillip G. Zaworski ◽  
Christian N. Parker
Keyword(s):  
Cell Wall ◽  
Cell Growth ◽  
High Throughput ◽  
High Throughput Screening ◽  
Osmotic Shock ◽  
Fungal Cell Wall ◽  
Cell Wall Synthesis ◽  
Fungal Cell ◽  
Unique Nature ◽  
Pharmacologically Active

Fungal cell wall synthesis is essential for viability, requiring the activity of genes involved in environmental sensing, precursor synthesis, transport, secretion, and assembly. This multitude of potential targets, the availability of known agents targeting this pathway, and the unique nature of fungal cell wall synthesis make this pathway an appealing target for drug discovery. Here we describe the adaptation of an assay monitoring cell wall synthesis for high-throughput screening. The assay requires fungal cell growth, in the presence of the test compound, for 3 h before the cells are subjected to osmotic shock in the presence of a dye that stains DNA. Miniaturization of the assay to a 384-well plate format and removing a mechanical transfer led to subtle changes in the assay characteristics. Validation of the assay with a library of known pharmacologically active agents has identified a number of different classes of compounds that are active in this assay, causing aberrant cell wall morphology and in many cases the inhibition of fungal cell growth.

A highly selective fluorescent probe for real-time imaging of bacterial NAT2 and high-throughput screening of natural inhibitors for tuberculosis therapy

2019 ◽  
Vol 3 (1) ◽  
pp. 145-150 ◽  
Author(s):  
Yinzhu Jin ◽  
Zhenhao Tian ◽  
Xiangge Tian ◽  
Lei Feng ◽  
Jingnan Cui ◽  
...  
Keyword(s):  
Cell Wall ◽  
Real Time ◽  
Fluorescent Probe ◽  
High Throughput ◽  
High Throughput Screening ◽  
Molecular Target ◽  
Cell Wall Synthesis ◽  
Key Enzyme ◽  
Tuberculosis Therapy ◽  
Natural Inhibitors

Fluorescent probeARHBis developed for detecting in various bacteria the activity ofN-acetyltransferase 2 (NAT2), a key enzyme in cell wall synthesis and widely considered to be a molecular target for anti-mycobacterial therapy.

scholarly journals Mycobacterium tuberculosis CwsA Interacts with CrgA and Wag31, and the CrgA-CwsA Complex Is Involved in Peptidoglycan Synthesis and Cell Shape Determination

2012 ◽  
Vol 194 (23) ◽  
pp. 6398-6409 ◽  
Author(s):  
P. Plocinski ◽  
N. Arora ◽  
K. Sarva ◽  
E. Blaszczyk ◽  
H. Qin ◽  
...  
Keyword(s):  
Cell Wall ◽  
Mycobacterium Tuberculosis ◽  
Mutant Strain ◽  
Cell Shape ◽  
Double Mutant ◽  
Bacterial Cell Division ◽  
Cell Wall Synthesis ◽  
Content Type ◽  
Ring Assembly ◽  
Double Mutant Strain

ABSTRACTBacterial cell division and cell wall synthesis are highly coordinated processes involving multiple proteins. Here, we show that Rv0008c, a novel small membrane protein fromMycobacterium tuberculosis, localizes to the poles and on membranes and shows an overall punctate localization throughout the cell. Furthermore, Rv0008c interacts with two proteins, CrgA and Wag31, implicated in peptidoglycan (PG) synthesis in mycobacteria. Deletion of the Rv0008c homolog inM. smegmatis, MSMEG_0023, caused bulged cell poles, formation of rounded cells, and defects in polar localization of Wag31 and cell wall synthesis, with cell wall synthesis measured by the incorporation of the [14C]N-acetylglucosamine cell wall precursor. TheM. smegmatisMSMEG_0023crgAdouble mutant strain showed severe defects in growth, viability, cell wall synthesis, cell shape, and the localization of the FtsZ, FtsI, and Wag31 proteins. The double mutant strain also exhibited increased autolytic activity in the presence of detergents. Because CrgA and Wag31 proteins interact with FtsI individually, we believe that regulated cell wall synthesis and cell shape maintenance require the concerted actions of the CrgA, Rv0008c, FtsI, and Wag31 proteins. We propose that, together, CrgA and Rv0008c, renamed CwsA forcellwall synthesis and cellshape proteinA, play crucial roles in septal and polar PG synthesis and help coordinate these processes with the FtsZ-ring assembly in mycobacteria.

Differential Assay for High-Throughput Screening of Antibacterial Compounds

2007 ◽  
Vol 12 (8) ◽  
pp. 1102-1108 ◽  
Author(s):  
Shaun P. Falk ◽  
Andrew T. Ulijasz ◽  
Bernard Weisblum
Keyword(s):  
Cell Wall ◽  
High Throughput ◽  
High Throughput Screening ◽  
Immediate Release ◽  
Cell Wall Synthesis ◽  
Compound Library ◽  
Fluorogenic Substrate ◽  
Cell Wall Disruption ◽  
Surfactant Activity ◽  
High Throughput Assay

The previously described Bacillus subtilis reporter strain BAU-102 is capable of detecting cell wall synthesis inhibitors that act at all stages of the cell wall synthesis pathway. In addition, this strain is capable of detecting compounds with hydrophobic/ surfactant activity and alternative mechanisms of cell wall disruption. BAU-102 sequesters preformed β-gal in the periplasm, suggesting leakage of β-gal as the means by which this assay detects compound activities. A model is proposed according to which β-gal release by BAU-102 reflects activation of pathways leading to autolysis. The authors also report a simplified high-throughput assay using BAU-102 combined with the fluorogenic substrate N-methylumbelliferyl-β-D-galactoside as a single reagent. Cell wall inhibitors release β-gal consistently only after 60 min of incubation, whereas compounds with surfactant activity show an almost immediate release. A high-throughput screen of a 480-compound library of known bioactives yielded 8 compounds that cause β-gal release. These results validate the BAU-102 assay as an effective tool in antimicrobial drug discovery. ( Journal of Biomolecular Screening 2007:1102-1108)

scholarly journals Cell-wall synthesis and ribosome maturation are co-regulated by an RNA switch in Mycobacterium tuberculosis

2018 ◽  
Vol 46 (11) ◽  
pp. 5837-5849 ◽  
Author(s):  
Stefan Schwenk ◽  
Alexandra Moores ◽  
Irene Nobeli ◽  
Timothy D McHugh ◽  
Kristine B Arnvig
Keyword(s):  
Cell Wall ◽  
Mycobacterium Tuberculosis ◽  
Cell Wall Synthesis ◽  
Rna Switch
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