Regulation Mechanism of the Lateral Diffusion of Band 3 in Erythrocyte Membranes by the Membrane Skeleton

Tomishige, Michio; Sako, Yasushi; Kusumi, Akihiro

doi:10.1083/jcb.142.4.989

Cited by 194 publications

(221 citation statements)

References 69 publications

(116 reference statements)

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“…Evidence for an interaction between specific membrane domains and the cytoskeleton in animal cells has repeatedly been reported (Kusumi and Sako, 1996;Rodgers and Zavzavadjian, 2001). Actin, spectrin and vimentin have been implicated in retarding the movement of membrane proteins in various mammalian cells (Sheetz et al, 1980;Tomishige et al, 1998;Oliferenko et al, 1999;Runembert et al, 2002). In yeast cells, this question has not been investigated.…”

Section: Discussionmentioning

confidence: 99%

Visualization of Protein Compartmentation within the Plasma Membrane of Living Yeast Cells

Malínská

Malínský

Opekarová

et al. 2003

MBoC

269

319

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Different distribution patterns of the arginine/H؉ symporter Can1p, the H ؉ plasma membrane ATPase Pma1p, and the hexose transport facilitator Hxt1p within the plasma membrane of living Saccharomyces cerevisiae cells were visualized using fluorescence protein tagging of these proteins. Although Hxt1p-GFP was evenly distributed through the whole cell surface, Can1p-GFP and Pma1p-GFP were confined to characteristic subregions in the plasma membrane. Pma1p is a well-documented raft protein. Evidence is presented that Can1p, but not Hxt1p, is exclusively associated with lipid rafts, too. Double labeling experiments with Can1p-GFP-and Pma1p-RFP-containing cells demonstrate that these proteins occupy two different nonoverlapping membrane microdomains. The size of Can1p-rich (Pma1p-poor) areas was estimated to 300 nm. These domains were shown to be stable in growing cells for >30 min. To our knowledge, this is the first observation of a cell polarization-independent lateral compartmentation in the plasma membrane of a living cell. INTRODUCTIONLipid rafts (detergent-resistant membranes, detergent-insoluble glycolipid-enriched microdomains, or glycolipid-enriched membranes) are dynamic assemblies enriched in sterols and sphingolipids occurring laterally distributed in the plasma membrane of most, if not all, eukaryotes (for reviews, see Simons and Ikonen, 1997;Brown and London, 1998). In mammalian cells cholesterol, sphingomyelin, and glycosphingolipids are the basic constituents of detergentresistant membranes (Rietveld and Simons, 1998), whereas in yeast these are ergosterol, inositolphosphoceramide, and its mannosylated derivatives (Kü bler et al., 1996;Bagnat et al., 2000). Existence of similar plasma membrane domains in plant cells can also be expected, although it has not been reported so far.A distinctive feature of lipid rafts is their insolubility in mild nonionic detergents (typically Triton X-100) at 4°C (Brown and Rose, 1992;Rietveld and Simons, 1998). Consequently, they are found floating in low-density fractions of solubilizates of mild detergent-treated membranes. Rafts obtained by this procedure selectively recruit specific membrane proteins, whereas others are excluded. The recognition of selective protein retention within the detergent-resistant domains led to a concept of lateral subcompartmentation within the plasma membrane (Simons and Ikonen, 1997;Brown and London, 1998).It was proposed that rafts form a platform for lipid (Simons and Ikonen, 2000) and protein sorting and trafficking (Simons and van Meer, 1988;Galbiati et al., 2001;Ikonen, 2001) and cell signaling (Field et al., 1997;Stauffer and Meyer, 1997;Simons and Toomre, 2000;Dykstra et al., 2001). Lipid rafts provide the cells also with a mechanism for functional and spatial control of exocytosis (Chamberlain et al., 2001) and are involved in immune cell activation (for reviews, see Dykstra et al., 2001;Galbiati et al., 2001;Katagiri et al., 2001). Accumulating evidence documents that compositionally distinct lipid microdomains are assembled ...

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Section: Discussionmentioning

confidence: 99%

Visualization of Protein Compartmentation within the Plasma Membrane of Living Yeast Cells

Malínská

Malínský

Opekarová

et al. 2003

MBoC

269

319

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“…The movies were recorded on a digital videotape (PDV-184ME; Sony). Quantitative analysis of ABCA1 movement was performed based on mean square displacement (MSD) methods described previously (39)(40)(41)(42)(43). For each trajectory of a particle, MSD, <(Δr(Δt)) 2 >, every time interval was calculated according to the equation (41,44) …”

Section: Methodsmentioning

confidence: 99%

ABCA1 dimer–monomer interconversion during HDL generation revealed by single-molecule imaging

Nagata

Nakada

Kasai

et al. 2013

Proc. Natl. Acad. Sci. U.S.A.

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The generation of high-density lipoprotein (HDL), one of the most critical events for preventing atherosclerosis, is mediated by ATPbinding cassette protein A1 (ABCA1). ABCA1 is known to transfer cellular cholesterol and phospholipids to apolipoprotein A-I (apoA-I) for generating discoidal HDL (dHDL) particles, composed of 100-200 lipid molecules surrounded by two apoA-I molecules; however, the regulatory mechanisms are still poorly understood. Here we observed ABCA1-GFP and apoA-I at the level of single molecules on the plasma membrane via a total internal reflection fluorescence microscope. We found that about 70% of total ABCA1-GFP spots are immobilized on the plasma membrane and estimated that about 89% of immobile ABCA1 molecules are in dimers. Furthermore, an ATPase-deficient ABCA1 mutant failed to be immobilized or form a dimer. We found that the lipid acceptor apoA-I interacts with the ABCA1 dimer to generate dHDL and is followed by ABCA1 dimermonomer interconversion. This indicates that the formation of the ABCA1 dimer is the key for apoA-I binding and nascent HDL generation. Our findings suggest the physiological significance of conversion of the ABCA1 monomer to a dimer: The dimer serves as a receptor for two apoA-I molecules for dHDL particle generation.membrane protein | transporter P lasma high-density lipoprotein (HDL) is critical for preventing coronary artery disease (1). A member of the ATPdependent transporter family of ABC proteins, ATP-binding cassette protein A1 (ABCA1) initiates the generation of discoidal HDL (dHDL), a bilayer fragment consisting of 100-200 lipids wrapped by two molecules of apolipoprotein A-I (apoA-I) (2-4), by exporting cholesterol and phospholipids to lipid-free apoA-I in serum (5). More than 70 mutations have been identified in the ABCA1 gene. Indeed, mutations in ABCA1 lead to Tangier disease, which is characterized by plasma HDL deficiency (6-10). ABCA1 has two large extracellular domains (ECDs), and the two intramolecular disulfide bonds between them are necessary for apoA-I binding and HDL formation (11-13) (Fig. 1A). Two pieces of evidence suggest that apoA-I interacts with a specific conformation of the ECDs in an ATP-dependent manner: Chemical cross-linkers can cross-link apoA-I with ABCA1, and ATPasedeficient ABCA1 mutants fail to mediate apoA-I binding and crosslinking. However, the importance of direct binding of ABCA1-apoA-I in HDL formation is still controversial and, furthermore, how a dHDL particle containing two molecules of apoA-I is formed from lipid-free apoA-I monomers and membrane lipids is unknown.To address these issues, we performed single-molecule fluorescence imaging (14, 15) of ABCA1 and apoA-I on the plasma membrane (PM) via a total internal reflection fluorescence (TIRF) microscope in living cells. We examined the dynamic behaviors of ABCA1 as well as the interaction of ABCA1 with apoA-I, and found that ABCA1 forms an immobile dimer on the PM; the ABCA1 dimer then dissociates into diffusing monomers upon interaction with apoA-I, finally gen...

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“…Another diffusion type was described according to these observations, called hop diffusion. It is claimed that both lipids and trans-membrane proteins diffuse in small compartments whose boundaries were mostly determined by cytoskeleton of the cell until they change the compartment by hop diffusion (Tomishige et al 1998). Although there are many studies claiming that saturated lipids and GPI-anchored proteins show confined diffusion on cell membrane on islands of 80 nm and 700 nm in size (Schutz et al 2000;Lenne et al 2006;Wenger et al 2007), some other SPT studies has recently claimed that the diffusion of saturated and unsaturated lipids is not different in cell membrane, which can be because of very small size (,16 nm) or very fast association/dissociation of the rafts.…”

Section: Applicationsmentioning

confidence: 99%

Fluorescence Techniques to Study Lipid Dynamics

Sezgin¹,

Schwille²

2011

Cold Spring Harbor Perspectives in Biology

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Biological research has always tremendously benefited from the development of key methodology. In fact, it was the advent of microscopy that shaped our understanding of cells as the fundamental units of life. Microscopic techniques are still central to the elucidation of biological units and processes, but equally important are methods that allow access to the dimension of time, to investigate the dynamics of molecular functions and interactions. Here, fluorescence spectroscopy with its sensitivity to access the single-molecule level, and its large temporal resolution, has been opening up fully new perspectives for cell biology. Here we summarize the key fluorescent techniques used to study cellular dynamics, with the focus on lipid and membrane systems.T o elucidate cellular processes in their native dynamic environment has been one of the main issues in cell biology over the past decades. The lack of appropriate techniques has long been the main limiting step for the research on dynamic systems, because it was impossible to acquire real time information with the well-known biochemical techniques. The key challenge in dynamically observing biological systems is to combine the ability to resolve moderate to very low concentrations of molecules-because they are simply limited in living cells-on relevant timescales. Relevant timescales in cell biology can be minutes and hours, on a systemic level of cell metabolism, down to the microsecond and even nanosecond regime in which molecular and intramolecular rearrangements take place. With respect to lipidic systems, relevant dynamics range from the local movements of lipids by diffusion to the mechanical transformations of whole membranes, spanning several orders of magnitude in time to be covered. Like for other cellular processes, the investigation of lipids and membranes also in general benefited greatly from the introduction of fluorescence microscopy and spectroscopy to biology. After the 1960s, great technological inventions based on the phenomenon of fluorescence were made, such as confocal microscopy, fluorescence recovery after photobleaching (FRAP), fluorescence correlation spectroscopy (FCS), Förster resonance energy transfer (FRET), total internal reflection fluorescence (TIRF), and two-photon microscopy, that not only revolutionized imaging but also yielded access to dynamics on previously inaccessible timescales. Another very big step was certainly taken after the introduction of fluorescent proteins, which again accelerated the use of these techniques in living cells and organisms. Nowadays, the technical advancements of fluorescence-based methods allow us to explore systems as small as single molecules with temporal resolution down to the nanoseconds regime. Lately, even the resolution limit of optical microscopy, for a long time being one of the fundamental barriers in elucidating cellular processes, has been overcome by smart applications of the phenomenon of fluorescence.This article aims at giving a short overview on mainly fluorescence-based metho...

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Regulation Mechanism of the Lateral Diffusion of Band 3 in Erythrocyte Membranes by the Membrane Skeleton

Cited by 194 publications

References 69 publications

Visualization of Protein Compartmentation within the Plasma Membrane of Living Yeast Cells

Visualization of Protein Compartmentation within the Plasma Membrane of Living Yeast Cells

ABCA1 dimer–monomer interconversion during HDL generation revealed by single-molecule imaging

Fluorescence Techniques to Study Lipid Dynamics

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