Phosphorylation of unlabeled proteins can be detected using immunological or enzymatic techniques. Anti-phosphotyrosine antibodies are used with immunoblots to detect tyrosine phosphorylation. This unit presents a protocol employing anti-phosphotyrosine antibodies with detection by either (125)I-labeled protein A or enhanced chemiluminescence (ECL).
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ROS-induced oxidative stress is a major contributor to sperm cryoinjury.
Hum Reprod
February 2024
Institute of Science and Technology, National Health Commission, Beijing, China.
Study Question: What is the mechanism behind cryoinjury in human sperm, particularly concerning the interplay between reactive oxygen species (ROS) and autophagy, and how does it subsequently affect sperm fate?
Summary Answer: The freeze-thaw operation induces oxidative stress by generating abundant ROS, which impairs sperm motility and activates autophagy, ultimately guiding the sperm toward programmed cell death such as apoptosis and necrosis, as well as triggering premature capacitation.
What Is Known Already: Both ROS-induced oxidative stress and autophagy are thought to exert an influence on the quality of frozen-thawed sperm.
Study Design, Size, Duration: Overall, 84 semen specimens were collected from young healthy fertile males, with careful quality evaluation.
Analysis of intracellular tyrosine phosphorylation in circulating neutrophils as a rapid assay for the effect of oral tyrosine kinase inhibitors.
Front Pharmacol
April 2023
Department of Physiology, Semmelweis University School of Medicine, Budapest, Hungary.
Tyrosine kinases are crucial signaling components of diverse biological processes and are major therapeutic targets in various malignancies and immune-mediated disorders. A critical step of development of novel tyrosine kinase inhibitors is the transition from the confirmation of the effects of drug candidates to the analysis of their efficacy. To facilitate this transition, we have developed a rapid assay for the analysis of the effect of oral tyrosine kinase inhibitors on basal tyrosine phosphorylation of circulating mouse neutrophils.
View Article and Find Full Text PDFPhosphoproteome profiling of hippocampal synaptic plasticity.
Biochem Biophys Res Commun
October 2022
Rare Disease Research Center, Korea Research Institute of Bioscience and Biotechnology, Daejeon, 34141, South Korea. Electronic address:
The balance between the actions of protein kinases and phosphatases is crucial for neuronal functions, including synaptic plasticity. Although the phosphorylation and dephosphorylation of neuronal proteins are regulated by synaptic plasticity, no systematic analyses of this have yet been conducted. We performed a phosphoproteomic analysis of hippocampal synaptic plasticity using a nano-Acquity/Synapt LC-MS/MS system.
View Article and Find Full Text PDFLow molecular weight protein phosphatase APH mediates tyrosine dephosphorylation and ABA response in Arabidopsis.
Stress Biol
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Shanghai Center for Plant Stress Biology, CAS Center for Excellence in Molecular Plant Sciences, Chinese Academy of Sciences, Shanghai, 200032 China.
Unlabelled: Low molecular weight protein tyrosine phosphatase (LWM-PTP), also known as acid phosphatase, is a highly conserved tyrosine phosphatase in living organisms. However, the function of LWM-PTP homolog has not been reported yet in plants. Here, we revealed a homolog of acid phosphatase, APH, in Arabidopsis plants, is a functional protein tyrosine phosphatase.
View Article and Find Full Text PDFEffects of Electrical Stimulation on the Signal Transduction-Related Proteins, c-Src and Focal Adhesion Kinase, in Fibroblasts.
Life (Basel)
April 2022
Laboratory of Human Anatomy and Cell Biology, Faculty of Health Sciences, Tsukuba University of Technology, Tsukuba 305-8521, Japan.
Electrical stimulation of the skin and muscles, e.g., in the fields of rehabilitation medicine and acupuncture, is known to locally increase blood flow and metabolism, and thus have beneficial health effects.
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