Protein stability influences many aspects of biology, and measuring their stability in vivo can provide important insights into biological systems.This chapter describes in detail two methods to assess the stability of a specific protein based on its transient expression in Arabidopsis protoplasts. First, a pulse-chase assay based on radioactive metabolic labeling of cellular proteins, followed by immunoprecipitation of the protein of interest. The decrease in radioactive signal is monitored over time and can be used to determine the protein's half-life.Alternatively, we also present a nonradioactive assay based on the use of reporter proteins, whose ratio can be quantified. This assay can be used to determine the relative stability of a protein of interest under specific conditions.
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http://dx.doi.org/10.1007/978-1-0716-2784-6_13 | DOI Listing |
Mol Pharm
January 2025
Department of Nuclear Medicine, Fudan University Shanghai Cancer Center; Department of Oncology, Shanghai Medical College, Fudan University, Shanghai 200032, China.
This study aimed to develop and evaluate a novel fibroblast activation protein (FAP)-specific tracer, fluorine-18-labeled fibroblast activation protein inhibitor-FUSCC-07 ([F]F-FAPI-FUSCC-07), for use in both preclinical and clinical settings. Preclinical evaluations were conducted to assess the stability and partition coefficient of [F]F-FAPI-FUSCC-07. Experiments involving human glioma U87MG cells demonstrated its cellular uptake and inhibitory properties.
View Article and Find Full Text PDFLife Metab
August 2022
College of Life Sciences, Hubei Key Laboratory of Cell Homeostasis, TaiKang Center for Life and Medical Sciences, TaiKang Medical School, Wuhan University, Wuhan, China.
Low-density lipoprotein (LDL) is the main carrier of cholesterol and cholesteryl ester in circulation. High plasma levels of LDL cholesterol (LDL-C) are a major risk factor of atherosclerotic cardiovascular disease (ASCVD). LDL-C lowering is recommended by many guidelines for the prevention and treatment of ASCVD.
View Article and Find Full Text PDFInnovation (Camb)
January 2025
AIM Center, College of Life Sciences and Technology, Beijing University of Chemical Technology, Institute of Microbiology, Chinese Academy of Sciences, Beijing, China.
Predicting free energy changes (ΔΔG) is essential for enhancing our understanding of protein evolution and plays a pivotal role in protein engineering and pharmaceutical development. While traditional methods offer valuable insights, they are often constrained by computational speed and reliance on biased training datasets. These constraints become particularly evident when aiming for accurate ΔΔG predictions across a diverse array of protein sequences.
View Article and Find Full Text PDFIn Silico Pharmacol
January 2025
Department of Biotechnology and Bioinformatics, JSS Academy of Higher Education and Research, Mysore, Karnataka 570015 India.
Unlabelled: Parkinson's Disease (PD) is a neurodegenerative disorder that primarily affects persons aged 65 and older. It leads to a decline in motor function as a result of the buildup of abnormal protein deposits called Lewy bodies in the brain. Existing therapies exhibit restricted effectiveness and undesirable side effects.
View Article and Find Full Text PDFIn Silico Pharmacol
January 2025
Teady Bioscience Research Laboratory, Ilara Mokin, Ondo State Nigeria.
Unlabelled: Breast cancer remains a global health challenge, with rising cases predicted in the coming decades. The complexity of breast cancer treatment arises from its complex nature, often involving multiple therapeutic strategies. One promising approach is targeting the ERK5 pathway, a key regulator in cancer cell proliferation and survival.
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