Peptidyl-tRNA hydrolase (Pth) activity ensures the rapid recycling of peptidyl-tRNAs that result from premature termination of translation. Historically, the hydrolyzing activity of Pth has been assayed with radiolabeled N-blocked aminoacyl-tRNAs in assay systems that require the separation of radiolabeled amino acid from the N-blocked aminoacyl-tRNA complex. In the present study, we describe the development of a kinetic fluorescence polarization (FP) assay that enables measurements of Pth activity without the need to separate bound and free tracer. The hydrolyzing activity of Pth was determined by measuring the change in polarization values that resulted from the cleavage of a fluorescently labeled substrate (BODIPY-Lys-tRNA(Lys)). The data were analyzed using an equation describing first-order dissociation and the results showed that the experimental data correlated well with the theoretical curve. A runs test of the residuals showed that the experimental data did not significantly differ from the first-order model. The assay is adaptable to a multiwell format and is sensitive enough to detect Pth-like activity in bacterial cell lysate. The Pth FP assay provides a homogeneous and kinetic format for measuring Pth activity in vitro.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1006/abio.2002.5700 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Characterization of jamun () juice fortified with nanoemulsified vitamin D: and assessment of its nutraceutical value and anti-diabetic potential.
Food Chem X
January 2025
Microbial and Pharmaceutical Biotechnology Laboratory, Department of Pharmacognosy and Phytochemistry, School of Pharmaceutical Education and Research, Jamia Hamdard, New Delhi, India.
This study aimed to fortify Jamun () juice with vitamin D to address vitamin D deficiency and boost health. A nanoemulsion of vitamin D was fabricated using a low-temperature (4-20C) sonication method and incorporated into the juice. The vitamin D fortified jamun juice (VDFJJ) exhibited a total polyphenol content of 14.
View Article and Find Full Text PDFGlial activation and nociceptive neuropeptide elevation associated with the development of chronic post-traumatic headache following repetitive blast exposure.
Neurobiol Pain
December 2024
Virginia Polytechnic Institute and State University. Department of Biomedical Engineering, 325 Stranger St., Blacksburg, VA 24060, United States.
Chronic headaches and pain are prevalent in those who are exposure to blast events, yet there is a gap in fundamental data that identifies the pathological mechanism for the chronification of pain. Blast-related post-traumatic headaches (PTH) are understudied and chronic pain behaviors in preclinical models can be vital to help elucidate PTH mechanisms. The descending pain modulatory system controls pain perception and involves specific brain regions such as the cortex, thalamus, pons, and medulla.
View Article and Find Full Text PDFFabricating Lattice-Confined Pt Single Atoms With High Electron-Deficient State for Alkali Hydrogen Evolution Under Industrial-Current Density.
Adv Mater
January 2025
State Key Laboratory of Organic-Inorganic Composites and College of Chemical Engineering, Beijing University of Chemical Technology, Beijing, 100029, People's Republic of China.
The confining effect is essential to regulate the activity and stability of single-atom catalysts (SACs), but the universal fabrication of confined SACs is still a great challenge. Here, various lattice-confined Pt SACs supported by different carriers are constructed by a universal co-reduction approach. Notably, Pt single atoms confined in the lattice of Ni(OH) (Pt/Ni(OH)) with a high electron-deficient state exhibit excellent activity for basic hydrogen evolution reaction (HER).
View Article and Find Full Text PDFPrimary Hyperparathyroidism: A Common Condition With an Uncommon Location.
Cureus
December 2024
Endocrinology Department, Hospital de Egas Moniz - Centro Hospitalar de Lisboa Ocidental, Lisbon, PRT.
Primary hyperparathyroidism (PHPT) is a prevalent clinical condition characterized by an inappropriate secretion of parathyroid hormone (PTH). It is most often caused by one or more parathyroid adenomas, which can, in rare cases, be ectopically located. Ectopic adenomas can pose a diagnostic challenge, lead to treatment delay, and be a common cause of recurrent hypercalcemia after parathyroidectomy.
View Article and Find Full Text PDFChemogenetic modulation of parathyroid hormone secretion alleviates osteoporosis in ovariectomized rats.
Biochem Biophys Res Commun
January 2025
Guangdong Provincial Key Laboratory of Brain Connectome and Behavior, Shenzhen-Hong Kong Institute of Brain Science, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, 518055, China; CAS Key Laboratory of Brain Connectome and Manipulation, The Brain Cognition and Brain Disease Institute, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen, 518055, China; University of Chinese Academy of Sciences, Beijing, 100049, China. Electronic address:
Parathyroid hormone (PTH) is critical for regulating calcium and phosphate homeostasis, and its dysregulation contributes to osteoporosis. Current methods for precise control of PTH secretion are limited. This study explores chemogenetic tools to regulate PTH secretion in parathyroid chief cells via Gq/Gi signaling.
View Article and Find Full Text PDFWant 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!