AI Article Synopsis
Article Abstract
Aberrations in protein modification with ubiquitin-fold modifier (UFM1) are associated with a range of diseases, but the biological function and regulation of this post-translational modification, known as UFMylation, remain enigmatic. To provide activity-based probes for UFMylation, we have developed a new method for the installation of electrophilic warheads at the C-terminus of recombinant UFM1. A C-terminal UFM1 acyl hydrazide was readily produced by selective intein cleavage and chemoselectively acylated by a variety of carboxylic acid anhydrides at pH 3, without detriment to the folded protein or reactions at unprotected amino acid side chains. The resulting UFM1 activity-based probes show a range of tunable reactivity and high selectivity for proteins involved in UFMylation processes; structurally related E1s, E2s, and proteases associated with Ub or other Ubls were unreactive. The UFM1 probes were active both in cell lysates and in living cells. A previously inaccessible α-chloroacetyl probe was remarkably selective for covalent modification of the active-site cysteine of de-UFMylase UFSP2 .
Download full-text PDF |
Source |
|---|---|
| http://www.ncbi.nlm.nih.gov/pmc/articles/PMC9228560 | PMC |
| http://dx.doi.org/10.1021/acscentsci.2c00203 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Age-related lung changes linked to altered lysosomal protease profile, histology, and ultrastructure.
PLoS One
December 2024
Division of Pharmacology and Toxicology, Department of Pharmaceutical Sciences, University of Vienna, Vienna, Austria.
Introduction: The aging process is intricately linked to alterations in cellular and tissue structures, with the respiratory system being particularly susceptible to age-related changes. Therefore, this study aimed to profile the activity of proteases using activity-based probes in lung tissues of old and young rats, focusing on the expression levels of different, in particular cathepsins G and X and matrix Metalloproteinases (MMPs). Additionally, the impact on extracellular matrix (ECM) components, particularly fibronectin, in relation to age-related histological and ultrastructural changes in lung tissues was investigated.
View Article and Find Full Text PDFScreening Traditional Chinese Medicine Compounds for Inhibiting UCHL3 Activity Based on Molecular Docking and Deubiquitinating Enzyme Probe Technology.
J Vis Exp
November 2024
Guanganmen Hospital, China Academy of Chinese Medical Sciences;
Deubiquitinating enzymes (DUBs) play a pivotal role in modulating ubiquitination homeostasis, with UCHL3 being an archetypal cysteine DUB intricately involved in a myriad of physiological and pathological processes. Therefore, developing small molecule inhibitors targeting Ubiquitin C-Terminal Hydrolase L3 (UCHL3) is of great significance. This protocol aims to establish a process for virtual screening and in vitro validation of small molecule inhibitors of cysteine DUB represented by UCHL3.
View Article and Find Full Text PDFDifferential behavioral engagement of inhibitory interneuron subtypes in the zebra finch brain.
Neuron
December 2024
NYU Neuroscience Institute and Department of Otolaryngology, New York University Langone Medical Center, New York, NY, USA; Center for Neural Science, New York University, New York, NY, USA. Electronic address:
Inhibitory interneurons are highly heterogeneous circuit elements often characterized by cell biological properties, but how these factors relate to specific roles underlying complex behavior remains poorly understood. Using chronic silicon probe recordings, we demonstrate that distinct interneuron groups perform different inhibitory roles within HVC, a song production circuit in the zebra finch forebrain. To link these functional subtypes to molecular identity, we performed two-photon targeted electrophysiological recordings of HVC interneurons followed by post hoc immunohistochemistry of subtype-specific markers.
View Article and Find Full Text PDFIdentification of TIGAR, a direct proteomic target associated with the hypoglycemic effect of Berberine.
Fitoterapia
December 2024
State Key Laboratory of Southwest Chinese Medicine Resources, School of Pharmacy, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, Sichuan, PR China. Electronic address:
Diabetes mellitus is a global chronic metabolic disease and the prevalence of diabetes mellitus is increasing dramatically every year. Berberine (BBR) from Coptidis Rhizoma has potent hypoglycemic effects, however, the specific proteins targeted by berberine that contribute to its hypoglycemic action remain to be elucidated. In this work, TIGAR (TP53-induced glycolysis and apoptosis regulator) was identified as a direct target protein for berberine using activity-based protein profiling (ABPP) and other chemical proteomics techniques with active photoaffinity probes as chemical tools.
View Article and Find Full Text PDF/ Value Determination for Penicillin-Binding Proteins in Live Cells.
ACS Infect Dis
December 2024
Department of Medicinal Chemistry, University of Minnesota, 208 Harvard Street SE, Minneapolis, Minnesota 55454, United States.
Article Synopsis
- - Penicillin-binding proteins (PBPs) are crucial bacterial enzymes that can be inhibited by β-lactam antibiotics, disrupting their ability to grow and divide, ultimately leading to cell lysis.
- - Traditional metrics like IC values for measuring enzyme inhibition can be misleading, especially for covalent inhibitors, making the second-order rate constant (k) a better measure of potency.
- - The researchers created a whole-cell k assay using a fluorescent probe to assess β-lactam potency across multiple PBPs, confirming its effectiveness in relation to existing IC values and supporting further structure-activity relationship studies.
Want AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!