Inactivation of NPC1L1 causes multiple lipid transport defects and protects against diet-induced hypercholesterolemia.
AuthorsDavies JP, Scott C, Oishi K, Liapis A, Ioannou YA
JournalJ Biol Chem
PubMed ID15671032
'NPC1L1, a recently identified relative of Niemann-Pick C1, was characterized to determine its subcellular location and potential function(s). NPC1L1 was highly expressed in HepG2 cells and localized in a subcellular vesicular compartment rich in the small GTPase Rab5. mRNA expression profiling revealed significant differences between mouse and man with highest ... More
Dynamics and function of phospholipase D and phosphatidic acid during phagocytosis.
AuthorsCorrotte M, Chasserot-Golaz S, Huang P, Du G, Ktistakis NT, Frohman MA, Vitale N, Bader MF, Grant NJ
JournalTraffic
PubMed ID16497229
'Phospholipase D (PLD) produces phosphatidic acid (PA), an established intracellular signalling lipid that has been also implicated in vesicular trafficking, and as such, PLD could play multiple roles during phagocytosis. Using an RNA interference strategy, we show that endogenous PLD1 and PLD2 are necessary for efficient phagocytosis in murine macrophages, ... More
Rab11-family interacting proteins define spatially and temporally distinct regions within the dynamic Rab11a-dependent recycling system.
AuthorsBaetz NW, Goldenring JR,
JournalMol Biol Cell
PubMed ID23283983
'The Rab11-family interacting proteins (Rab11-FIPs) facilitate Rab11-dependent vesicle recycling. We hypothesized that Rab11-FIPs define discrete subdomains and carry out temporally distinct roles within the recycling system. We used live-cell deconvolution microscopy of HeLa cells expressing chimeric fluorescent Rab11-FIPs to examine Rab11-FIP localization, transferrin passage through Rab11-FIP-containing compartments, and overlap among ... More
Site-specific protein labeling by Sfp phosphopantetheinyl transferase.
AuthorsYin J, Lin AJ, Golan DE, Walsh CT,
JournalNat Protoc
PubMed ID17406245
'Sfp phosphopantetheinyl transferase covalently attaches small-molecule probes including biotin and various organic fluorophores to a specific serine residue in the peptidyl carrier protein (PCP) or a short 11-residue peptide tag ybbR through a phosphopantetheinyl linker. We describe here a protocol for site-specific protein labeling by Sfp-catalyzed protein post-translational modification that ... More
Correlative light-electron microscopy (CLEM) combining live-cell imaging and immunolabeling of ultrathin cryosections.
'The visualization of fluorescent proteins in living cells is a powerful approach to study intracellular dynamics. A limitation of fluorescence imaging, however, is that it lacks fine structural information; a fluorescent spot could represent an entire organelle, an organellar subdomain or even aggregates of proteins or membranes. These limitations can ... More
Wide-field subdiffraction imaging by accumulated binding of diffusing probes.
AuthorsSharonov A, Hochstrasser RM
JournalProc Natl Acad Sci U S A
PubMed ID17142314
A method is introduced for subdiffraction imaging that accumulates points by collisional flux. It is based on targeting the surface of objects by fluorescent probes diffusing in the solution. Because the flux of probes at the object is essentially constant over long time periods, the examination of an almost unlimited ... More
Single-cell FRET imaging of transferrin receptor trafficking dynamics by Sfp-catalyzed, site-specific protein labeling.
AuthorsYin J, Lin AJ, Buckett PD, Wessling-Resnick M, Golan DE, Walsh CT,
JournalChem Biol
PubMed ID16183024
Fluorescence imaging of living cells depends on an efficient and specific method for labeling the target cellular protein with fluorophores. Here we show that Sfp phosphopantetheinyl transferase-catalyzed protein labeling is suitable for fluorescence imaging of membrane proteins that spend at least part of their membrane trafficking cycle at the cell ... More
Mechanism for amyloid precursor-like protein 2 enhancement of major histocompatibility complex class I molecule degradation.
AuthorsTuli A, Sharma M, Capek HL, Naslavsky N, Caplan S, Solheim JC,
JournalJ Biol Chem
PubMed ID19808674
Earlier studies have demonstrated interaction of the murine major histocompatibility complex (MHC) class I molecule K(d) with amyloid precursor-like protein 2 (APLP2), a ubiquitously expressed member of the amyloid precursor protein family. Our current findings indicate that APLP2 is internalized in a clathrin-dependent manner, as shown by utilization of inhibitors ... More
Dynamin 2 mutations associated with human diseases impair clathrin-mediated receptor endocytosis.
AuthorsBitoun M, Durieux AC, Prudhon B, Bevilacqua JA, Herledan A, Sakanyan V, Urtizberea A, Cartier L, Romero NB, Guicheney P,
JournalHum Mutat
PubMed ID19623537
Dynamin 2 (DNM2) is a large GTPase involved in the release of nascent vesicles during endocytosis and intracellular membrane trafficking. Distinct DNM2 mutations, affecting the middle domain (MD) and the Pleckstrin homology domain (PH), have been identified in autosomal dominant centronuclear myopathy (CNM) and in the intermediate and axonal forms ... More
Genetically encoded short peptide tags for orthogonal protein labeling by Sfp and AcpS phosphopantetheinyl transferases.
AuthorsZhou Z, Cironi P, Lin AJ, Xu Y, Hrvatin S, Golan DE, Silver PA, Walsh CT, Yin J
JournalACS Chem Biol
PubMed ID17465518
Short peptide tags S6 and A1, each 12 residues in length, were identified from a phage-displayed peptide library as efficient substrates for site-specific protein labeling catalyzed by Sfp and AcpS phosphopantetheinyl transferases (PPTases), respectively. S6 and A1 tags were selected for useful levels of orthogonality in reactivities with the PPTases: ... More
Regulation of ubiquitin-binding proteins by monoubiquitination.
AuthorsHoeller D, Crosetto N, Blagoev B, Raiborg C, Tikkanen R, Wagner S, Kowanetz K, Breitling R, Mann M, Stenmark H, Dikic I
JournalNat Cell Biol
PubMed ID16429130
Proteins containing ubiquitin-binding domains (UBDs) interact with ubiquitinated targets and regulate diverse biological processes, including endocytosis, signal transduction, transcription and DNA repair. Many of the UBD-containing proteins are also themselves monoubiquitinated, but the functional role and the mechanisms that underlie this modification are less well understood. Here, we demonstrate that ... More
Plasma membrane domains specialized for clathrin-mediated endocytosis in primary cells.
AuthorsBellve KD, Leonard D, Standley C, Lifshitz LM, Tuft RA, Hayakawa A, Corvera S, Fogarty KE
JournalJ Biol Chem
PubMed ID16537543
Clathrin assembly at the plasma membrane is a fundamental process required for endocytosis. In cultured cells, most of the clathrin is localized to large patches that display little lateral mobility. The functional role of these regions is not clear, and it has been thought that they may represent artifacts of ... More
Vectorial insertion of apical and basolateral membrane proteins in polarized epithelial cells revealed by quantitative 3D live cell imaging.
AuthorsHua W, Sheff D, Toomre D, Mellman I
JournalJ Cell Biol
PubMed ID16567501
Although epithelial cells are known to exhibit a polarized distribution of membrane components, the pathways responsible for delivering membrane proteins to their appropriate domains remain unclear. Using an optimized approach to three-dimensional live cell imaging, we have visualized the transport of newly synthesized apical and basolateral membrane proteins in fully ... More
Dynamin deficiency causes insulin secretion failure and hyperglycemia.
Authors
JournalProc Natl Acad Sci U S A
PubMed ID34362840
Dynamin 2 regulates biphasic insulin secretion and plasma glucose homeostasis.
Authors
JournalJ Clin Invest
PubMed ID26413867
ADVANCED IMAGING. Extended-resolution structured illumination imaging of endocytic and cytoskeletal dynamics.
Authors
JournalScience
PubMed ID26315442
Imaging vesicle formation dynamics supports the flexible model of clathrin-mediated endocytosis.
Authors
JournalNat Commun
PubMed ID35365614
Dynamin regulates the dynamics and mechanical strength of the actin cytoskeleton as a multifilament actin-bundling protein.
Authors
JournalNat Cell Biol
PubMed ID32451441
Cargo-specific recruitment in clathrin- and dynamin-independent endocytosis.
Authors
JournalNat Cell Biol
PubMed ID34616024
TDP-43 Depletion in Microglia Promotes Amyloid Clearance but Also Induces Synapse Loss.
Authors
JournalNeuron
PubMed ID28669544
SNAREs define targeting specificity of trafficking vesicles by combinatorial interaction with tethering factors.
Authors
JournalNat Commun
PubMed ID30962439
Lipid-gated monovalent ion fluxes regulate endocytic traffic and support immune surveillance.