Cellular heterogeneity occurs, and should be probed, at multiple levels of cellular structure and physiology from the genome to enzyme activity. In particular, single-cell measures of protein levels are complemented by single-cell measurements of the activity of these proteins. Microfluidic assays of enzyme activity at the single-cell level combine moderate to high throughput with low dead volumes and the potential for automation. Herein, we describe the steps required to fabricate and operate a microfluidic device for chemical cytometry of fluorescent or fluorogenic reporters of enzyme activity in individual cells.
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http://dx.doi.org/10.1007/978-1-4939-2987-0_15 | DOI Listing |
Sci Rep
December 2024
Department of Life Sciences, Pohang University of Science and Technology, Pohang, 37673, Kyungbook, Republic of Korea.
Alanine racemase (Alr) catalyzes the pyridoxal 5'-phosphate (PLP)-dependent racemization between L- and D-alanine in bacteria. Owing to the potential interest in targeting Alr for antibacterial drug development, several studies have determined the structures of Alr from different species, proposing models for the reaction mechanism. Insights into its reaction dynamics may be conducive to a better understanding of the Alr reaction mechanism.
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December 2024
Promega Corporation, 2800 Woods Hollow Road, Madison, WI, 53711, USA.
The cyclic GMP-AMP synthase-stimulator of the interferon gene (cGAS-STING) signaling pathway is considered an essential pattern recognition and effector pathway in the natural immune system and is mainly responsible for recognizing DNA molecules present in the cytoplasm and activating downstream signaling pathways to generate type I interferons (IFN-I) and other inflammatory factors. STING, a crucial junction protein in the innate immune system, exerts an essential role in host resistance to external pathogen invasion. The DNA introduced by pathogens or tumors is recognized by the cytoplasmic nucleic acid receptor cGAS, and a second messenger, cGAMP, is generated using intracellular guanosine triphosphate (GTP) and adenosine triphosphate (ATP).
View Article and Find Full Text PDFBMC Infect Dis
December 2024
Department of Community Health Sciences, Aga Khan University, Karachi, Pakistan.
Background: Mucocutaneous leishmaniasis (MCL) is a severe form of leishmaniasis causing chronic and destructive lesions. Accurate diagnosis is crucial for effective treatment. Traditional methods, such as the Montenegro skin test is delayed hypersensitivity test.
View Article and Find Full Text PDFBMC Microbiol
December 2024
Department of Microbiology, Faculty of Medicine, Shahed University, Tehran, Iran.
Background: Klebsiella pneumoniae is a clinically relevant pathogen that has raised considerable public health concerns. This study aims to determine the presence of beta-lactamase genes and perform molecular genotyping of multidrug-resistant (MDR) K. pneumoniae clinical isolates.
View Article and Find Full Text PDFJ Biomol Struct Dyn
December 2024
Department of Bioinformatics, School of Life Sciences Pondicherry University, Puducherry, India.
Flavin adenine nucleotide (FAD)-dependent oxidoreductase enzyme Alcohol oxidase (AOX) facilitates the growth of methylotrophic yeast C. boidinii by catabolizing methanol, producing formaldehyde and hydrogen peroxide. Vacuolar Protease-A (PrA) from C.
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