Clathrin-mediated endocytosis manages the vesicular transport of the bulk of membrane proteins from the plasma membrane and the trans-Golgi network. During this process, discrete sets of adaptor proteins recognize specific classes of membrane proteins, which recruit and assemble clathrin lattices on the membrane. An important determinant to the success of this vesicular transport reaction is the intrinsic ability of adaptors to polymerize clathrin on a membrane surface. Adaptor-induced clathrin assembly has traditionally been analyzed using static electron microscopy-based approaches. Here, we describe a methodology to follow adaptor-induced clathrin assembly in real-time using fluorescence microscopy on a facile model membrane assay system of supported membrane tubes (SMrT). Results from such assays can be conveniently run through routine image analysis procedures to extract kinetic parameters of the clathrin assembly reaction.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1007/978-1-4939-8719-1_12 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
A new statistical metric for robust target detection in cryo-EM using 2D template matching.
IUCrJ
March 2025
RNA Therapeutics Institute, University of Massachusetts Chan Medical School, Worcester, USA.
2D template matching (2DTM) can be used to detect molecules and their assemblies in cellular cryo-EM images with high positional and orientational accuracy. While 2DTM successfully detects spherical targets such as large ribosomal subunits, challenges remain in detecting smaller and more aspherical targets in various environments. In this work, a novel 2DTM metric, referred to as the 2DTM p-value, is developed to extend the 2DTM framework to more complex applications.
View Article and Find Full Text PDFThe nanoscale organization of the Nipah virus fusion protein informs new membrane fusion mechanisms.
Elife
January 2025
Institute of Parasitology, Faculty of Agricultural and Environmental Sciences, McGill University, Montreal, Canada.
Paramyxovirus membrane fusion requires an attachment protein for receptor binding and a fusion protein for membrane fusion triggering. Nipah virus (NiV) attachment protein (G) binds to ephrinB2 or -B3 receptors, and fusion protein (F) mediates membrane fusion. NiV-F is a class I fusion protein and is activated by endosomal cleavage.
View Article and Find Full Text PDFFast-diffusing receptor collisions with slow-diffusing peptide ligand assemble the ternary parathyroid hormone-GPCR-arrestin complex.
Nat Commun
December 2024
Department of Pharmacology and Chemical Biology, University of Pittsburgh School of Medicine, Pittsburgh, PA, 15261, USA.
Article Synopsis
- - The study investigates how a peptide hormone (PTH) interacts with its receptor (PTHR) and β-arrestin (βarr) to form a ternary complex, which is key for G protein-coupled receptor (GPCR) signaling.
- - Using fluorescent markers and advanced imaging techniques, the research shows that PTHR moves freely in the cell membrane while unbound PTH has limited mobility, indicating a distinct dynamic behavior.
- - The formation of the PTH-PTHR-βarr complex happens in three steps: ligand-receptor collisions, βarr recruitment triggered by a specific lipid (PIP), and final assembly within clathrin clusters, highlighting the importance of PIP in GPCR
Haploinsufficiency and Alzheimer's Disease: The Possible Pathogenic and Protective Genetic Factors.
Int J Mol Sci
November 2024
Department of Bionano Technology, Gachon Medical Research Institute, Gachon University, Seongnam 13120, Republic of Korea.
Alzheimer's disease (AD) is a complex neurodegenerative disorder influenced by various genetic factors. In addition to the well-established amyloid precursor protein (), Presenilin-1 (), Presenilin-2 (), and apolipoprotein E (), several other genes such as Sortilin-related receptor 1 (), Phospholipid-transporting ATPase ABCA7 (), Triggering Receptor Expressed on Myeloid Cells 2 (), Phosphatidylinositol-binding clathrin assembly protein (), and clusterin () were implicated. These genes contribute to neurodegeneration through both gain-of-function and loss-of-function mechanisms.
View Article and Find Full Text PDFIntersectin1 promotes clathrin-mediated endocytosis by organizing and stabilizing endocytic protein interaction networks.
Cell Rep
December 2024
Department of Molecular and Cell Biology, University of California, Berkeley, Berkeley, CA 94720, USA. Electronic address:
During clathrin-mediated endocytosis (CME), dozens of proteins are recruited to nascent CME sites on the plasma membrane, and their spatial and temporal coordination is crucial for efficient CME. Here, we show that the scaffold protein intersectin1 (ITSN1) promotes CME by organizing and stabilizing endocytic protein interaction networks. Live-cell imaging of genome-edited cells revealed that endogenously labeled ITSN1 is recruited during CME site stabilization and growth and that ITSN1 knockdown impairs endocytic protein recruitment during this stage.
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!