Protocol to visualize and quantify the COPII concentration and anterograde transport of nascent G protein-coupled receptors.

STAR Protoc

Department of Pharmacology and Toxicology, Medical College of Georgia, Augusta University, Augusta, GA 30912, USA. Electronic address:

Published: June 2024

Here, we present a protocol for visualization and quantification of the recruitment of newly synthesized G protein-coupled receptors (GPCRs) to coat protein complex II vesicles and GPCR transport from the endoplasmic reticulum through the Golgi to the cell surface in the retention using the selective hooks assay. We describe steps for plasmid construction, cell transfection, transport synchronization, confocal microscope imaging, and quantification. This protocol is also applicable for studying the transport of non-GPCR cargoes. For complete details on the use and execution of this protocol, please refer to Xu et al..

Download full-text PDF

Source
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC10951583PMC
http://dx.doi.org/10.1016/j.xpro.2024.102955DOI Listing

Publication Analysis

Top Keywords

protein-coupled receptors
8
protocol
4
protocol visualize
4
visualize quantify
4
quantify copii
4
copii concentration
4
concentration anterograde
4
transport
4
anterograde transport
4
transport nascent
4

Similar Publications

Cancer pain is one of the most common symptoms in patients with advanced cancer. In this study, we aimed to investigate the effects of the -related gene C (MrgC) receptors on bone cancer pain. Mechanical withdrawal threshold (MWT) and thermal withdrawal latency (TWL) were measured after the inoculation of Walker 256 mammary gland carcinoma cells into the tibia of adult Sprague-Dawley rats.

View Article and Find Full Text PDF

Skin, as the body's largest organ, acts as the primary defense mechanism against infection and injury. The maintenance of skin health heavily relies on the regulation of epidermal stem cells, crucial for ensuring epidermal homeostasis, hair regeneration, and the repair of epidermal injuries. Recent studies have placed a growing emphasis on G protein-coupled receptor (GPCR) in the context of understanding epidermal stem cells, uncovering its significant role in determining their fate.

View Article and Find Full Text PDF

Osteoarthritis (OA) is a degenerative joint disease that affects the cartilage and surrounding tissues. The transcription factor Kruppel-like family factor 9 (KLF9) has been identified as a regulator of tumorigenesis. However, its role in OA is still not fully understood.

View Article and Find Full Text PDF

Unsymmetric triazine-based triglucoside detergents for membrane protein stability.

Chembiochem

January 2025

Hanyang University, Department of Bionano Engineering, 55 Hanyangdaehak-ro, 15588, Ansan, KOREA, REPUBLIC OF.

Membrane proteins play a crucial role in a variety of biological processes and are key targets for pharmaceutical development. Structural studies of membrane proteins provide molecular insights into the mechanisms of these processes and are essential for effective drug discovery. Historically, these studies have relied on solubilization of the target protein using detergents, but conventional detergents often fail to maintain the stability of challenging membrane proteins.

View Article and Find Full Text PDF

Split Membrane: A New Model to Accelerate All-Atom MD Simulation of Phospholipid Bilayers.

J Chem Inf Model

January 2025

CEITEC─Central European Institute of Technology, Masaryk University, Kamenice 753/5, 625 00 Brno, Czech Republic.

All-atom molecular dynamics simulations are powerful tools for studying cell membranes and their interactions with proteins and other molecules. However, these processes occur on time scales determined by the diffusion rate of phospholipids, which are challenging to achieve in all-atom models. Here, we present a new all-atom model that accelerates lipid diffusion by splitting phospholipid molecules into head and tail groups.

View Article and Find Full Text PDF

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!