Podosome-Type Adhesions and Focal Adhesions, So Alike Yet So Different Block Marc R
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Membrane Tension Buffering by Caveolae: a Role in Cancer?
Cancer and Metastasis Reviews (2020) 39:505–517 https://doi.org/10.1007/s10555-020-09899-2 Membrane tension buffering by caveolae: a role in cancer? Vibha Singh1 & Christophe Lamaze1 Published online: 30 May 2020 # Springer Science+Business Media, LLC, part of Springer Nature 2020 Abstract Caveolae are bulb-like invaginations made up of two essential structural proteins, caveolin-1 and cavins, which are abundantly present at the plasma membrane of vertebrate cells. Since their discovery more than 60 years ago, the function of caveolae has been mired in controversy. The last decade has seen the characterization of new caveolae components and regulators together with the discovery of additional cellular functions that have shed new light on these enigmatic structures. Early on, caveolae and/ or caveolin-1 have been involved in the regulation of several parameters associated with cancer progression such as cell migration, metastasis, angiogenesis, or cell growth. These studies have revealed that caveolin-1 and more recently cavin-1 have a dual role with either a negative or a positive effect on most of these parameters. The recent discovery that caveolae can act as mechanosensors has sparked an array of new studies that have addressed the mechanobiology of caveolae in various cellular functions. This review summarizes the current knowledge on caveolae and their role in cancer development through their activity in membrane tension buffering. We propose that the role of caveolae in cancer has to be revisited through their response to the mechanical forces encountered by cancer cells during tumor mass development. Keywords Caveolae . Cancer . Mechanosensing . Mechanotransdcution . Membrane tension . -
How Does Protein Zero Assemble Compact Myelin?
Preprints (www.preprints.org) | NOT PEER-REVIEWED | Posted: 13 May 2020 doi:10.20944/preprints202005.0222.v1 Peer-reviewed version available at Cells 2020, 9, 1832; doi:10.3390/cells9081832 Perspective How Does Protein Zero Assemble Compact Myelin? Arne Raasakka 1,* and Petri Kursula 1,2 1 Department of Biomedicine, University of Bergen, Jonas Lies vei 91, NO-5009 Bergen, Norway 2 Faculty of Biochemistry and Molecular Medicine & Biocenter Oulu, University of Oulu, Aapistie 7A, FI-90220 Oulu, Finland; [email protected] * Correspondence: [email protected] Abstract: Myelin protein zero (P0), a type I transmembrane protein, is the most abundant protein in peripheral nervous system (PNS) myelin – the lipid-rich, periodic structure that concentrically encloses long axonal segments. Schwann cells, the myelinating glia of the PNS, express P0 throughout their development until the formation of mature myelin. In the intramyelinic compartment, the immunoglobulin-like domain of P0 bridges apposing membranes together via homophilic adhesion, forming a dense, macroscopic ultrastructure known as the intraperiod line. The C-terminal tail of P0 adheres apposing membranes together in the narrow cytoplasmic compartment of compact myelin, much like myelin basic protein (MBP). In mouse models, the absence of P0, unlike that of MBP or P2, severely disturbs the formation of myelin. Therefore, P0 is the executive molecule of PNS myelin maturation. How and when is P0 trafficked and modified to enable myelin compaction, and how disease mutations that give rise to incurable peripheral neuropathies alter the function of P0, are currently open questions. The potential mechanisms of P0 function in myelination are discussed, providing a foundation for the understanding of mature myelin development and how it derails in peripheral neuropathies. -
Structural Insights Into Membrane Fusion Mediated by Convergent Small Fusogens
cells Review Structural Insights into Membrane Fusion Mediated by Convergent Small Fusogens Yiming Yang * and Nandini Nagarajan Margam Department of Microbiology and Immunology, Dalhousie University, Halifax, NS B3H 4R2, Canada; [email protected] * Correspondence: [email protected] Abstract: From lifeless viral particles to complex multicellular organisms, membrane fusion is inarguably the important fundamental biological phenomena. Sitting at the heart of membrane fusion are protein mediators known as fusogens. Despite the extensive functional and structural characterization of these proteins in recent years, scientists are still grappling with the fundamental mechanisms underlying membrane fusion. From an evolutionary perspective, fusogens follow divergent evolutionary principles in that they are functionally independent and do not share any sequence identity; however, they possess structural similarity, raising the possibility that membrane fusion is mediated by essential motifs ubiquitous to all. In this review, we particularly emphasize structural characteristics of small-molecular-weight fusogens in the hope of uncovering the most fundamental aspects mediating membrane–membrane interactions. By identifying and elucidating fusion-dependent functional domains, this review paves the way for future research exploring novel fusogens in health and disease. Keywords: fusogen; SNARE; FAST; atlastin; spanin; myomaker; myomerger; membrane fusion 1. Introduction Citation: Yang, Y.; Margam, N.N. Structural Insights into Membrane Membrane fusion -
Sphingosine-1-Phosphate Receptor 2 Controls Podosome Components Induced by RANKL Affecting Osteoclastogenesis and Bone Resorption
cells Article Sphingosine-1-Phosphate Receptor 2 Controls Podosome Components Induced by RANKL Affecting Osteoclastogenesis and Bone Resorption Li-Chien Hsu 1, Sakamuri V. Reddy 2, Özlem Yilmaz 1 and Hong Yu 1,* 1 Department of Oral Health Sciences, College of Dental Medicine, Medical University of South Carolina, Charleston, SC 29425, USA; [email protected] (L.-C.H.); [email protected] (Ö.Y.) 2 Department of Pediatrics, Osteoclast Center, Darby Children’s Research Institute, Medical University of South Carolina, Charleston, SC 29425, USA; [email protected] * Correspondence: [email protected]; Tel.: +1-843-792-0635 Received: 17 November 2018; Accepted: 26 December 2018; Published: 1 January 2019 Abstract: Proinflammatory cytokine production, cell chemotaxis, and osteoclastogenesis can lead to inflammatory bone loss. Previously, we showed that sphingosine-1-phosphate receptor 2 (S1PR2), a G protein coupled receptor, regulates inflammatory cytokine production and osteoclastogenesis. However, the signaling pathways regulated by S1PR2 in modulating inflammatory bone loss have not been elucidated. Herein, we demonstrated that inhibition of S1PR2 by a specific S1PR2 antagonist (JTE013) suppressed phosphoinositide 3-kinase (PI3K), mitogen-activated protein kinases (MAPKs), and nuclear factor kappa-B (NF-κB) induced by an oral bacterial pathogen, Aggregatibacter actinomycetemcomitans, and inhibited the release of IL-1β, IL-6, TNF-α, and S1P in murine bone marrow cells. In addition, shRNA knockdown of S1PR2 or treatment by JTE013 suppressed cell chemotaxis induced by bacteria-stimulated cell culture media. Furthermore, JTE013 suppressed osteoclastogenesis and bone resorption induced by RANKL in murine bone marrow cultures. ShRNA knockdown of S1PR2 or inhibition of S1PR2 by JTE013 suppressed podosome components, including PI3K, Src, Pyk2, integrin β3, filamentous actin (F-actin), and paxillin levels induced by RANKL in murine bone marrow cells. -
A Novel Role for Calpain 4 in Podosome Assembly
A NOVEL ROLE FOR CALPAIN 4 IN PODOSOME ASSEMBLY by Thomas Riley Dowler A thesis submitted to the Department of Biochemistry In conformity with the requirements for the degree of Masters of Science Queen’s University Kingston, Ontario, Canada September, 2008 Copyright © Thomas Riley Dowler, 2008 Abstract Podosomes are adhesive and invasive structures which may play an important role in numerous physiological and pathological conditions including angiogenesis, atherosclerosis, and cancer metastasis. Recently, the cysteine protease m-calpain (m- Capn) has been shown to cleave cortactin, an integral component of the podosomal F- actin core, as well as various proteins found in the peripheral adhesive region leading to the disassembly of these dynamic structures. In this study, I investigated whether Capn plays a role in the formation of podosomes downstream of c-Src. I show that: 1) phorbol- 12, 13-dibutyrate (PDBu) as well as c-Src-Y527F expression induces podosome formation in mouse embryonic fibroblasts; 2) PDBu- and constitutively active c-Src- induced podosome formation is inhibited by the knockout of the m- and µ-Capn small regulatory subunit Capn4 in mouse embryonic fibroblasts (Capn4-/-), but is partially restored by re-expression of Capn4; 3) Capn4 localizes to podosomes; and 4) Inhibition of m- and µ-Capn proteolytic activity by the cell permeable calpain inhibitors has little effect on the formation of podosomes downstream of active c-Src. I conclude that Capn4 may play a role in the assembly phase of podosomes independent of calpain proteolytic activity. Work done in collaboration to determine a possible mechanism of action for the role of Capn4 in podosome assembly indicates that a possible binding partner of Capn4, β-PIX, co-localizes with, and shows in vivo association with Capn4. -
Lipoprotein Cholesterols Are Stored in High-Resistant
Preprints (www.preprints.org) | NOT PEER-REVIEWED | Posted: 10 October 2018 doi:10.20944/preprints201810.0211.v1 Lipoprotein cholesterols are stored in high-resistant, metastatic cancer cells and released upon stress: implication for a mechanism underlying hypocholesterolemia in cancer patients Taka Eguchi1,2,3,*, Chiharu Sogawa1,3, Kisho Ono1, Mami Itagaki1,3, Masaki Matsumoto1 1Department of Dental Pharmacology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Okayama, Japan. 2Advanced Research Center for Oral and Craniofacial Sciences, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, Okayama, Japan. 3Okayama University Dental School, Okayama, Japan. 4Department of Molecular and Cellular Biology, Medical Institute of Bioregulation, Kyushu University, 3-1-1 Maidashi, Higashi-ku, Fukuoka 812-8582, Japan. *Correspondence and requests for materials should be addressed to: Taka Eguchi, D.D.S., Ph.D. 2-5-1 Shikata-cho, Okayama 700-8525 Japan Phone: +81-86-235-6662; Fax: +81-86-235-6664 E-mail: [email protected] Author Contributions: TE conceptualized and designed the study. TE, CS, and MM prepared resources. TE, MM, CS, KO devised methodology. CS, MM, KO, MI carried out the experimentation. TE, KO, CS, MM interpreted data. TE wrote the manuscript. KO, CS revised and edited the manuscript. All authors reviewed the manuscript. 1 © 2018 by the author(s). Distributed under a Creative Commons CC BY license. Preprints (www.preprints.org) | NOT PEER-REVIEWED | Posted: 10 October 2018 doi:10.20944/preprints201810.0211.v1 Abstract Resistant cancer often shows a particular secretory trait such as heat shock proteins (HSPs) and extracellular vesicles (EVs), including exosomes and oncosomes surrounded by lipid bilayers. -
Therapeutic Nanobodies Targeting Cell Plasma Membrane Transport Proteins: a High-Risk/High-Gain Endeavor
biomolecules Review Therapeutic Nanobodies Targeting Cell Plasma Membrane Transport Proteins: A High-Risk/High-Gain Endeavor Raf Van Campenhout 1 , Serge Muyldermans 2 , Mathieu Vinken 1,†, Nick Devoogdt 3,† and Timo W.M. De Groof 3,*,† 1 Department of In Vitro Toxicology and Dermato-Cosmetology, Vrije Universiteit Brussel, Laarbeeklaan 103, 1090 Brussels, Belgium; [email protected] (R.V.C.); [email protected] (M.V.) 2 Laboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, Pleinlaan 2, 1050 Brussels, Belgium; [email protected] 3 In Vivo Cellular and Molecular Imaging Laboratory, Vrije Universiteit Brussel, Laarbeeklaan 103, 1090 Brussels, Belgium; [email protected] * Correspondence: [email protected]; Tel.: +32-2-6291980 † These authors share equal seniorship. Abstract: Cell plasma membrane proteins are considered as gatekeepers of the cell and play a major role in regulating various processes. Transport proteins constitute a subclass of cell plasma membrane proteins enabling the exchange of molecules and ions between the extracellular environment and the cytosol. A plethora of human pathologies are associated with the altered expression or dysfunction of cell plasma membrane transport proteins, making them interesting therapeutic drug targets. However, the search for therapeutics is challenging, since many drug candidates targeting cell plasma membrane proteins fail in (pre)clinical testing due to inadequate selectivity, specificity, potency or stability. These latter characteristics are met by nanobodies, which potentially renders them eligible therapeutics targeting cell plasma membrane proteins. Therefore, a therapeutic nanobody-based strategy seems a valid approach to target and modulate the activity of cell plasma membrane Citation: Van Campenhout, R.; transport proteins. -
Poji: a Fiji-Based Tool for Analysis of Podosomes and Associated Proteins Robert Herzog1, Koen Van Den Dries2, Pasquale Cervero1 and Stefan Linder1,*
© 2020. Published by The Company of Biologists Ltd | Journal of Cell Science (2020) 133, jcs238964. doi:10.1242/jcs.238964 TOOLS AND RESOURCES Poji: a Fiji-based tool for analysis of podosomes and associated proteins Robert Herzog1, Koen van den Dries2, Pasquale Cervero1 and Stefan Linder1,* ABSTRACT structure that contains actomyosin contractility regulators such as Podosomes are actin-based adhesion and invasion structures in a lymphocyte-specific protein 1 (LSP1) (Cervero et al., 2018) and variety of cell types, with podosome-forming cells displaying up to supervillin (Bhuwania et al., 2012). Podosomes display a remarkably several hundreds of these structures. Podosome number, distribution broad repertoire of functions, ranging from cell-matrix adhesion and and composition can be affected by experimental treatments or during extracellular matrix degradation, to antigen presentation and regular turnover, necessitating a tool that is able to detect even subtle mechanosensing, and we refer to several reviews that cover these differences in podosomal properties. Here, we present a Fiji-based various aspects (Albiges-Rizo et al., 2009; Alonso et al., 2019; macro code termed ‘Poji’ (‘podosome analysis by Fiji’), which serves as Destaing et al., 2011; Linder, 2007; Linder and Wiesner, 2015; an easy-to-use tool to characterize a variety of cellular and podosomal Paterson and Courtneidge, 2018; van den Dries et al., 2014, 2019). parameters, including area, fluorescence intensity, relative enrichment Of note, especially myeloid cells such as macrophages often form of associated proteins and radial podosome intensity profiles. This tool >500 podosomes per cell, making podosomes a prominent should be useful to gain more detailed insight into the regulation, component of the actin cytoskeleton of these cells. -
Immuno-Electron and Confocal Laser Scanning Microscopy of the Glycocalyx
biology Article Immuno-Electron and Confocal Laser Scanning Microscopy of the Glycocalyx Shailey Gale Twamley 1,2, Anke Stach 1, Heike Heilmann 3, Berit Söhl-Kielczynski 4, Verena Stangl 1,2, Antje Ludwig 1,2,*,† and Agnieszka Münster-Wandowski 3,*,† 1 Medizinische Klinik für Kardiologie und Angiologie, Charité—Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, 10117 Berlin, Germany; [email protected] (S.G.T.); [email protected] (A.S.); [email protected] (V.S.) 2 DZHK (German Centre for Cardiovascular Research), Partner Site, 10117 Berlin, Germany 3 Institute of Integrative Neuroanatomy, Charité—Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, 10117 Berlin, Germany; [email protected] 4 Institute for Integrative Neurophysiology—Universitätsmedizin Berlin, Corporate Member of Freie Universität Berlin, Humboldt-Universität zu Berlin, and Berlin Institute of Health, 10117 Berlin, Germany; [email protected] * Correspondence: [email protected] (A.L.); [email protected] (A.M.-W.) † These authors equally contributed. Simple Summary: The glycocalyx (GCX) is a hydrated, gel-like layer of biological macromolecules attached to the cell membrane. The GCX acts as a barrier and regulates the entry of external substances into the cell. The function of the GCX is highly dependent on its structure and composition. Citation: Twamley, S.G.; Stach, A.; Pathogenic factors can affect the protective structure of the GCX. We know very little about the three- Heilmann, H.; Söhl-Kielczynski, B.; dimensional organization of the GXC. The tiny and delicate structures of the GCX are difficult Stangl, V.; Ludwig, A.; to study by microscopic techniques. -
In This Issue
By Jane Bradbury In this issue Samp1 makes nuclear moves FOXO3, ROS and apoptosis Diagrams of eukaryotic cells often show the nucleus in the Forkhead box O (FOXO) transcription factors induce centre of the cell, but the position of the nucleus actually apoptosis and regulate the production of reactive oxygen changes during certain developmental stages and cellular species (ROS). In neuronal tumour cells derived from events. For example, during the polarisation that precedes advanced stage neuroblastoma, FOXO3 – the most fibroblast migration, the nucleus moves away from the prevalent FOXO in neuronal cells – is inactivated. future leading edge of the cell. This nuclear movement involves nesprin-2G (a Notably, however, chemotherapeutic agents can activate FOXO3 and nuclear envelope spectrin repeat protein) and the SUN-domain protein SUN2, induce apoptosis in neuronal tumour cells. Judith Hagenbuchner and which are components of the linker of nucleoskeleton and cytoskeleton (LINC) colleagues (p. 1191), who have been investigating the molecular events that complex that form transmembrane actin-associated nuclear (TAN) lines at the underlie FOXO3-induced apoptosis, now report that, in two neuroblastoma nuclear envelope. Now, on page 1099, Edgar Gomes and colleagues report that cell lines, FOXO3 represses mitochondrial respiration and induces cellular the nuclear envelope protein spindle-associated membrane protein 1 (Samp1, ROS in response to chemotherapy. They show that etoposide- and also known as NET5 and TMEM201) is also required for nuclear movement in doxorubicin-induced elevation of cellular ROS depends on FOXO3 migrating fibroblasts. By using siRNA knockdown, the authors show that activation and on induction of its transcriptional target Bim (BCL2L11), a Samp1 is involved in centrosome orientation and nuclear movement during pro-apoptotic protein. -
Membrane Structure
Membranes Chapter 5 Membrane Structure The fluid mosaic model of membrane structure contends that membranes consist of: -phospholipids arranged in a bilayer -globular proteins inserted in the lipid bilayer 2 3 1 Membrane Structure Cellular membranes have 4 components: 1. phospholipid bilayer 2. transmembrane proteins 3. interior protein network 4. cell surface markers 4 5 Phospholipids Phospholipid structure consists of -glycerol – a 3-carbon polyalcohol acting as a backbone for the phospholipid -2 fatty acids attached to the glycerol -phosphate group attached to the glycerol 6 2 Phospholipids The fatty acids are nonpolar chains of carbon and hydrogen. -Their nonpolar nature makes them hydrophobic (“water-fearing”). The phosphate group is polar and hydrophilic (“water-loving”). 7 Phospholipids The partially hydrophilic, partially hydrophobic phospholipid spontaneously forms a bilayer: -fatty acids are on the inside -phosphate groups are on both surfaces of the bilayer 8 9 3 Phospholipids Phospholipid bilayers are fluid. -hydrogen bonding of water holds the 2 layers together -individual phospholipids and unanchored proteins can move through the membrane -saturated fatty acids make the membrane less fluid than unsaturated fatty acids -warm temperatures make the membrane more fluid than cold temperatures 10 Membrane Proteins Membrane proteins have various functions: 1. transporters 2. enzymes 3. cell surface receptors 4. cell surface identity markers 5. cell-to-cell adhesion proteins 6. attachments to the cytoskeleton 11 12 4 Membrane Proteins -
Membrane Protein Structure Determination and Characterisation by Solution and Solid-State NMR
biology Review Membrane Protein Structure Determination and Characterisation by Solution and Solid-State NMR Vivien Yeh , Alice Goode and Boyan B. Bonev * School of Life Sciences, University of Nottingham, Nottingham NG7 2UH, UK; [email protected] (V.Y.); [email protected] (A.G.) * Correspondence: [email protected] Received: 21 October 2020; Accepted: 11 November 2020; Published: 12 November 2020 Simple Summary: Cells, life’s smallest units, are defined within the enclosure of thin, continuous membranes, which confine the molecular machinery required for the life and replication of cells. Crucially, membranes of cells establish and actively maintain distinctly different environments inside cells, including electrical and solute gradients vital to normal cellular functions. Membrane proteins are in charge of transport, electrical polarisation, signalling, membrane remodelling and other important functions. As such, membrane proteins are key drug targets and understanding their structure and function is essential to drug development and cellular control. Membrane proteins have physical characteristics that make such studies very challenging. Nuclear magnetic resonance is one advanced tool that enables structural studies of membrane proteins and their interactions at the atomic level of detail. We discuss the applications of NMR in solution and solid state to membrane protein studies alongside new developments in signal and sensitivity enhancement through dynamic nuclear polarisation. Abstract: Biological membranes define the interface of life and its basic unit, the cell. Membrane proteins play key roles in membrane functions, yet their structure and mechanisms remain poorly understood. Breakthroughs in crystallography and electron microscopy have invigorated structural analysis while failing to characterise key functional interactions with lipids, small molecules and membrane modulators, as well as their conformational polymorphism and dynamics.