AI Article Synopsis
- The chapter explains the process of expressing and purifying myosin, a type of cytoskeletal molecular motor, using an insect cell system.
- It details methods for characterizing the quality of the purified myosin through techniques like mass photometry and negative-stain electron microscopy (EM).
- Lastly, it describes how to conduct in vitro assays to observe how fluorescently labeled myosin moves along actin tracks and mentions adapting these assays for MINFLUX imaging.
Article Abstract
To understand the mechanics and kinetic properties of cytoskeletal molecular motors such as myosin, typically the motor of interest needs to be expressed and purified and then analyzed using a range of in vitro-based assays. In this chapter, we describe how to express and purify myosin using the insect cell system, how to characterize the purified protein by mass photometry and negative-stain EM to assess its quality, and how to perform in vitro assays in which fluorescently labeled myosin walks along actin tracks, including a brief description of adapting these assays for MINFLUX imaging.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1007/978-1-0716-4280-1_9 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Recombinant Porcine FGF1 Promotes Muscle Stem Cell Proliferation and Mitochondrial Function for Cultured Meat Production.
J Agric Food Chem
January 2025
Department of Traditional Chinese Medicine, Sir Run Run Shaw Hospital, School of Medicine, Zhejiang University, Hangzhou 310016, China.
Cultured meat is an emerging technology with the potential to meet future protein demands while addressing the challenges associated with traditional livestock farming. The production of cultured meat requires efficient, animal component-free systems for muscle stem cell (MuSC) expansion. Fibroblast growth factor 1 (FGF1) is a critical growth factor that regulates the MuSC function.
View Article and Find Full Text PDFEngineered Therapeutic Bacteria with High-Yield Membrane Vesicle Production Inspired by Eukaryotic Membrane Curvature for Treating Inflammatory Bowel Disease.
ACS Nano
January 2025
Department of Chemical Engineering, University of Waterloo, 200 University Avenue West, Waterloo, Ontario N2L 3G1, Canada.
Bacterial membrane vesicles (BMVs) are emerging as powerful natural nanoparticles with transformative potential in medicine and industry. Despite their promise, scaling up BMV production and ensuring stable isolation and storage remain formidable challenges that limit their broader application. Inspired by eukaryotic mechanisms of membrane curvature, we engineered DH5α to serve as a high-efficiency BMV factory.
View Article and Find Full Text PDFDetection of Hepatitis C Virus Infection from Patient Sera in Cell Culture Using Semi-Automated Image Analysis.
Viruses
November 2024
Department of Infectious Diseases, Molecular Virology, Section Virus-Host Interactions, Heidelberg University, 69120 Heidelberg, Germany.
The study of hepatitis C virus (HCV) replication in cell culture is mainly based on cloned viral isolates requiring adaptation for efficient replication in Huh7 hepatoma cells. The analysis of wild-type (WT) isolates was enabled by the expression of SEC14L2 and by inhibitors targeting deleterious host factors. Here, we aimed to optimize cell culture models to allow infection with HCV from patient sera.
View Article and Find Full Text PDFIsolation and Characterization of a Lytic Phage PaTJ Against .
Viruses
November 2024
Key Laboratory of Tropical Marine Bio-Resources and Ecology, South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou 510000, China.
is a major global threat to human health, and phage therapy has emerged as a promising strategy for treating infections caused by multidrug-resistant pathogens. In this study, we isolated and characterized a lytic phage, PaTJ, from wastewater. PaTJ belongs to the phage family , and is featured by short latency (30 min) and large burst size (10 PFU per infected cell).
View Article and Find Full Text PDFImplementation of Sensitive Method for Determination of Benzophenone and Camphor UV Filters in Human Urine.
Toxics
November 2024
Department of Food Analysis and Nutrition, Faculty of Food and Biochemical Technology, University of Chemistry and Technology, Prague, Technicka 3, 166 28 Prague, Czech Republic.
The level of the human body's burden of benzophenone and camphor ultraviolet (UV) filters can be estimated from their urinary levels. The present study describes the implementations and validation of the sensitive analytical method for the analysis of seven benzophenone and two camphor UV filters in urine. Sample preparation includes overnight enzymatic hydrolysis and ethyl acetate extraction followed by purification by dispersive solid-phase extraction using a sorbent Z-Sep.
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!