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..
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http://dx.doi.org/10.1016/j.xpro.2024.102955 | DOI Listing |
Sheng Li Xue Bao
December 2024
College of Life Sciences, Fujian Normal University; Fujian Key Laboratory of Developmental and Neuro Biology, Fuzhou 350117, China.
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 PDFSheng Li Xue Bao
December 2024
Skin Disease Research Institute, the Second Affiliated Hospital Zhejiang University School of Medicine, Hangzhou 310058, China.
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 PDFCommun Biol
January 2025
Department of Orthopedics, Shengjing Hospital of China Medical University, Shenyang, China.
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 PDFChembiochem
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 PDFJ 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.
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