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Reconstituted lipid vesicles

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https://www.readbyqxmd.com/read/29108957/matrix-vesicles-from-chondrocytes-and-osteoblasts-their-biogenesis-properties-functions-and-biomimetic-models
#1
REVIEW
Massimo Bottini, Saida Mebarek, Karen L Anderson, Agnieszka Strzelecka-Kiliszek, Lukasz Bozycki, Ana Maria Sper Simão, Maytê Bolean, Pietro Ciancaglini, Joanna Bandorowicz Pikula, Slawomir Pikula, David Magne, Niels Volkmann, Dorit Hanein, José Luis Millán, Rene Buchet
BACKGROUND: Matrix vesicles (MVs) are released from hypertrophic chondrocytes and from mature osteoblasts, the cells responsible for endochondral and membranous ossification. Under pathological conditions, they can also be released from cells of non-skeletal tissues such as vascular smooth muscle cells. MVs are extracellular vesicles of approximately 100-300nm diameter harboring the biochemical machinery needed to induce mineralization. SCOPE OF THE REVIEW: The review comprehensively delineates our current knowledge of MV biology and highlights open questions aiming to stimulate further research...
November 3, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/29106973/reconstitution-of-snare-proteins-into-solid-supported-lipid-bilayer-stacks-and-x-ray-structure-analysis
#2
Yihui Xu, Jan Kuhlmann, Martha Brennich, Karlo Komorowski, Reinhard Jahn, Claudia Steinem, Tim Salditt
SNAREs are known as an important family of proteins mediating vesicle fusion. For various biophysical studies, they have been reconstituted into supported single bilayers via proteoliposome adsorption and rupture. In this study we extended this method to the reconstitution of SNAREs into supported multilamellar lipid membranes, i.e. oriented multibilayer stacks, as an ideal model system for x-ray structure analysis (x-ray reflectivity and diffraction). The reconstitution was implemented through a pathway of proteomicelle, proteoliposome and multibilayer...
October 26, 2017: Biochimica et Biophysica Acta
https://www.readbyqxmd.com/read/29073046/nonequilibrium-fluctuations-of-lipid-membranes-by-the-rotating-motor-protein-f1f0-atp-synthase
#3
Víctor G Almendro-Vedia, Paolo Natale, Michael Mell, Stephanie Bonneau, Francisco Monroy, Frederic Joubert, Iván López-Montero
ATP synthase is a rotating membrane protein that synthesizes ATP through proton-pumping activity across the membrane. To unveil the mechanical impact of this molecular active pump on the bending properties of its lipid environment, we have functionally reconstituted the ATP synthase in giant unilamellar vesicles and tracked the membrane fluctuations by means of flickering spectroscopy. We find that ATP synthase rotates at a frequency of about 20 Hz, promoting large nonequilibrium deformations at discrete hot spots in lipid vesicles and thus inducing an overall membrane softening...
October 24, 2017: Proceedings of the National Academy of Sciences of the United States of America
https://www.readbyqxmd.com/read/29066949/regulation-of-exocytotic-fusion-pores-by-snare-protein-transmembrane-domains
#4
REVIEW
Zhenyong Wu, Sathish Thiyagarajan, Ben O'Shaughnessy, Erdem Karatekin
Calcium-triggered exocytotic release of neurotransmitters and hormones from neurons and neuroendocrine cells underlies neuronal communication, motor activity and endocrine functions. The core of the neuronal exocytotic machinery is composed of soluble N-ethyl maleimide sensitive factor attachment protein receptors (SNAREs). Formation of complexes between vesicle-attached v- and plasma-membrane anchored t-SNAREs in a highly regulated fashion brings the membranes into close apposition. Small, soluble proteins called Complexins (Cpx) and calcium-sensing Synaptotagmins cooperate to block fusion at low resting calcium concentrations, but trigger release upon calcium increase...
2017: Frontiers in Molecular Neuroscience
https://www.readbyqxmd.com/read/29058192/patch-clamp-recordings-of-the-kcsa-k-channel-in-unilamellar-blisters
#5
Kimberly Matulef, Francis I Valiyaveetil
Patch-clamp electrophysiology is the standard technique used for the high-resolution functional measurements on ion channels. While studies using patch clamp are commonly carried out following ion channel expression in a heterologous system such as Xenopus oocytes or tissue culture cells, these studies can also be carried out using ion channels reconstituted into lipid vesicles. In this chapter, we describe the methodology for reconstituting ion channels into liposomes and the procedure for the generation of unilamellar blisters from these liposomes that are suitable for patch clamp...
2018: Methods in Molecular Biology
https://www.readbyqxmd.com/read/29020444/biologically-complex-planar-cell-plasma-membranes-supported-on-polyelectrolyte-cushions-enhance-transmembrane-protein-mobility-and-retain-native-orientation
#6
Han-Yuan Liu, Wei-Liang Chen, Christopher K Ober, Susan Daniel
Reconstituted supported lipid bilayers (SLB) are widely used as in vitro cell-surface models because they are compatible with a variety of surface-based analytical techniques. However, one of the challenges of using SLBs as a model of the cell surface is the limited complexity in membrane composition, including the incorporation of transmembrane proteins and lipid diversity that may impact the activity of those proteins. Additionally, it is challenging to preserve the transmembrane protein native orientation, function, and mobility in SLBs...
October 24, 2017: Langmuir: the ACS Journal of Surfaces and Colloids
https://www.readbyqxmd.com/read/29019793/tuning-biomimetic-membrane-barrier-properties-by-hydrocarbon-cholesterol-and-polymeric-additives
#7
Marta Espina Palanco, Nils Skovgaard, Jesper Søndergaard Hansen, Kirstine Berg-Sorensen, Claus Helix-Nielsen
The barrier properties of cellular membranes are increasingly attracting attention as a source of inspiration for designing biomimetic membranes. The broad range of potential technological applications makes the use of lipid and lately also polymeric materials a popular choice for constructing biomimetic membranes, where the barrier properties can be controlled by the composition of the membrane constituent elements. Here we investigate the membrane properties reported by the light-induced proton pumping activity of bacteriorhodopsin (bR) reconstituted in three vesicle systems of different membrane composition...
October 11, 2017: Bioinspiration & Biomimetics
https://www.readbyqxmd.com/read/28958150/synthesis-and-characterization-of-tethered-lipid-assemblies-for-membrane-protein-reconstitution-review
#8
Rémi Veneziano, Claire Rossi, Alexandre Chenal, Catherine Brenner, Daniel Ladant, Joël Chopineau
Biological membranes and their related molecular mechanisms are essential for all living organisms. Membranes host numerous proteins and are responsible for the exchange of molecules and ions, cell signaling, and cell compartmentation. Indeed, the plasma membrane delimits the intracellular compartment from the extracellular environment and intracellular membranes. Biological membranes also play a major role in metabolism regulation and cellular physiology (e.g., mitochondrial membranes). The elaboration of membrane based biomimetic systems allows us to reconstitute and investigate, in controlled conditions, biological events occurring at the membrane interface...
September 28, 2017: Biointerphases
https://www.readbyqxmd.com/read/28930438/supported-planar-mammalian-membranes-as-models-of-in-vivo-cell-surface-architectures
#9
Han-Yuan Liu, Hannah Grant, Hung-Lun Hsu, Raya Sorkin, Filip Bošković, Gijs Wuite, Susan Daniel
Emerging technologies use cell plasma membrane vesicles or "blebs" as an intermediate to form molecularly complete, planar cell surface mimetics that are compatible with a variety of characterization tools and microscopy methods. This approach enables direct incorporation of membrane proteins into supported lipid bilayers without using detergents and reconstitution and preserves native lipids and membrane species. Such a system can be advantageous as in vitro models of in vivo cell surfaces for study of the roles of membrane proteins as drug targets in drug delivery, host-pathogen interactions, tissue engineering, and many other bioanalytical and sensing applications...
October 18, 2017: ACS Applied Materials & Interfaces
https://www.readbyqxmd.com/read/28912594/quantification-of-aquaporin-z-reconstituted-into-vesicles-for-biomimetic-membrane-fabrication
#10
Hui Xian Gan, Hu Zhou, Qingsong Lin, Yen Wah Tong
Aquaporin incorporated biomimetic membranes are anticipated to offer unprecedented desalination capabilities. However, the lack of accurate methods to quantify the reconstituted aquaporin presents a huge hurdle in investigating aquaporin performance and optimizing membrane fabrication. Herein, we present three quantification methods to determine the Aquaporin-Z reconstituted into E. coli lipid vesicles: 1) nanogold labeling with transmission electron microscopy (TEM) visualization, 2) nickel labeling with inductively coupled plasma-mass spectrometry (ICP-MS) and 3) gel electrophoresis...
September 14, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28858288/simultaneous-lipid-and-content-mixing-assays-for-in-vitro-reconstitution-studies-of-synaptic-vesicle-fusion
#11
Xiaoxia Liu, Alpay Burak Seven, Junjie Xu, Victoria Esser, Lijing Su, Cong Ma, Josep Rizo
This protocol describes reconstitution assays to study how the neurotransmitter release machinery triggers Ca(2+)-dependent synaptic vesicle fusion. The assays monitor fusion between proteoliposomes containing the synaptic vesicle SNARE synaptobrevin (with or without the Ca(2+) sensor synaptotagmin-1) and proteoliposomes initially containing the plasma membrane SNAREs syntaxin-1 and soluble NSF attachment protein (SNAP)-25. Lipid mixing (from fluorescence de-quenching of Marina-Blue-labeled lipids) and content mixing (from development of fluorescence resonance energy transfer (FRET) between phycoerythrin-biotin (PhycoE-Biotin) and Cy5-streptavidin trapped in the two proteoliposome populations) are measured simultaneously to ensure that true, nonleaky membrane fusion is monitored...
September 2017: Nature Protocols
https://www.readbyqxmd.com/read/28854360/spatiotemporal-control-of-lipid-conversion-actin-based-mechanical-forces-and-curvature-sensors-during-clathrin-ap-1-coated-vesicle-biogenesis
#12
Mihaela Anitei, Christoph Stange, Cornelia Czupalla, Christian Niehage, Kai Schuhmann, Pia Sala, Aleksander Czogalla, Theresia Pursche, Ünal Coskun, Andrej Shevchenko, Bernard Hoflack
Clathrin/adaptor protein-1-coated carriers connect the secretory and the endocytic pathways. Carrier biogenesis relies on distinct protein networks changing membrane shape at the trans-Golgi network, each regulating coat assembly, F-actin-based mechanical forces, or the biophysical properties of lipid bilayers. How these different hubs are spatiotemporally coordinated remains largely unknown. Using in vitro reconstitution systems, quantitative proteomics, and lipidomics, as well as in vivo cell-based assays, we characterize the protein networks controlling membrane lipid composition, membrane shape, and carrier scission...
August 29, 2017: Cell Reports
https://www.readbyqxmd.com/read/28842688/light-independent-phospholipid-scramblase-activity-of-bacteriorhodopsin-from-halobacterium-salinarum
#13
Alice Verchère, Wei-Lin Ou, Birgit Ploier, Takefumi Morizumi, Michael A Goren, Peter Bütikofer, Oliver P Ernst, George Khelashvili, Anant K Menon
The retinylidene protein bacteriorhodopsin (BR) is a heptahelical light-dependent proton pump found in the purple membrane of the archaeon Halobacterium salinarum. We now show that when reconstituted into large unilamellar vesicles, purified BR trimers exhibit light-independent lipid scramblase activity, thereby facilitating transbilayer exchange of phospholipids between the leaflets of the vesicle membrane at a rate >10,000 per trimer per second. This activity is comparable to that of recently described scramblases including bovine rhodopsin and fungal TMEM16 proteins...
August 25, 2017: Scientific Reports
https://www.readbyqxmd.com/read/28827281/munc18a-clusters-snare-bearing-liposomes-prior-to-trans-snare-zippering
#14
Matthew Grant Arnold, Pratikshya Adhikari, Baobin Kang, Hao Xu 徐昊
Sec1-Munc18 (SM) proteins co-operate with SNAREs {SNAP [soluble NSF (N-ethylmaleimide-sensitive factor) attachment protein] receptors} to mediate membrane fusion in eukaryotic cells. Studies of Munc18a/Munc18-1/Stxbp1 in neurotransmission suggest that SM proteins accelerate fusion kinetics primarily by activating the partially zippered trans-SNARE complex. However, accumulating evidence has argued for additional roles for SM proteins in earlier steps in the fusion cascade. Here, we investigate the function of Munc18a in reconstituted exocytic reactions mediated by neuronal and non-neuronal SNAREs...
September 24, 2017: Biochemical Journal
https://www.readbyqxmd.com/read/28822127/in-vitro-analysis-of-metabolite-transport-proteins
#15
Marc-Sven Roell, Franziska Kuhnert, Shirin Zamani-Nour, Andreas P M Weber
The photorespiratory cycle is distributed over four cellular compartments, the chloroplast, peroxisomes, cytoplasm, and mitochondria. Shuttling of photorespiratory intermediates between these compartments is essential to maintain the function of photorespiration. Specific transport proteins mediate the transport across biological membranes and represent important components of the cellular metabolism. Although significant progress was made in the last years on identifying and characterizing new transport proteins, the overall picture of intracellular metabolite transporters is still rather incomplete...
2017: Methods in Molecular Biology
https://www.readbyqxmd.com/read/28768431/adefovir-dipivoxil-loaded-proliposomal-powders-with-improved-hepatoprotective-activity-formulation-optimization-pharmacokinetic-and-biodistribution-studies
#16
Ghada A Abdelbary, Maha M Amin, Mohamed Y Zakaria, Sally A El Awdan
The present study aimed to prepare proliposomal formulae for improving the oral bioavailability of Adefovir Dipivoxil (AD), a nucleoside reverse transcriptase inhibitor effective against Hepatitis B virus (HBV). The prepared proliposomal formulae were characterized for entrapment efficiency (E.E.%), vesicle size and in-vitro drug release after reconstitution to conventional liposomes. The optimized formula (F9) with a maximum desirability value of 0.858 was selected having E.E.% of 71±3.3% with an average vesicle size of 164...
August 2, 2017: Journal of Liposome Research
https://www.readbyqxmd.com/read/28703789/capturing-suboptical-dynamic-structures-in-lipid-bilayer-patches-formed-from-free-standing-giant-unilamellar-vesicles
#17
Tripta Bhatia, Flemming Cornelius, John H Ipsen
There is accumulating evidence that the small-scale lateral organization of biological membranes has a crucial role in signaling and trafficking in cells. However, it has been difficult to characterize these features with existing methods for preparing and analyzing freestanding membranes, because the dynamics occurs below the optical resolution possible with these protocols. We have developed a protocol that permits the imaging of lipid nanodomains and lateral protein organization in membranes of giant unilamellar vesicles (GUVs)...
August 2017: Nature Protocols
https://www.readbyqxmd.com/read/28697898/fusion-domains-guide-the-oriented-insertion-of-light-driven-proton-pumps-into-liposomes
#18
Noah Ritzmann, Johannes Thoma, Stephan Hirschi, David Kalbermatter, Dimitrios Fotiadis, Daniel J Müller
One major objective of synthetic biology is the bottom-up assembly of minimalistic nanocells consisting of lipid or polymer vesicles as architectural scaffolds and of membrane and soluble proteins as functional elements. However, there is no reliable method to orient membrane proteins reconstituted into vesicles. Here, we introduce a simple approach to orient the insertion of the light-driven proton pump proteorhodopsin (PR) into liposomes. To this end, we engineered red or green fluorescent proteins to the N- or C-terminus of PR, respectively...
September 19, 2017: Biophysical Journal
https://www.readbyqxmd.com/read/28695207/peripheral-myelin-protein-22-alters-membrane-architecture
#19
Kathleen F Mittendorf, Justin T Marinko, Cheri M Hampton, Zunlong Ke, Arina Hadziselimovic, Jonathan P Schlebach, Cheryl L Law, Jun Li, Elizabeth R Wright, Charles R Sanders, Melanie D Ohi
Peripheral myelin protein 22 (PMP22) is highly expressed in myelinating Schwann cells of the peripheral nervous system. PMP22 genetic alterations cause the most common forms of Charcot-Marie-Tooth disease (CMTD), which is characterized by severe dysmyelination in the peripheral nerves. However, the functions of PMP22 in Schwann cell membranes remain unclear. We demonstrate that reconstitution of purified PMP22 into lipid vesicles results in the formation of compressed and cylindrically wrapped protein-lipid vesicles that share common organizational traits with compact myelin of peripheral nerves in vivo...
July 2017: Science Advances
https://www.readbyqxmd.com/read/28688734/first-moves-towards-photoautotrophic-synthetic-cells-in-vitro-study-of-photosynthetic-reaction-centre-and-cytochrome-bc1-complex-interactions
#20
Emiliano Altamura, Rosa Fiorentino, Francesco Milano, Massimo Trotta, Gerardo Palazzo, Pasquale Stano, Fabio Mavelli
Following a bottom-up synthetic biology approach it is shown that vesicle-based cell-like systems (shortly "synthetic cells") can be designed and assembled to perform specific function (for biotechnological applications) and for studies in the origin-of-life field. We recently focused on the construction of synthetic cells capable to converting light into chemical energy. Here we first present our approach, which has been realized so far by the reconstitution of photosynthetic reaction centre in the membrane of giant lipid vesicles...
October 2017: Biophysical Chemistry
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