scholarly journals ER membranes exhibit phase behavior at sites of organelle contact

2020 ◽  
Vol 117 (13) ◽  
pp. 7225-7235 ◽  
Author(s):  
Christopher King ◽  
Prabuddha Sengupta ◽  
Arnold Y. Seo ◽  
Jennifer Lippincott-Schwartz
Keyword(s):  
Phase Behavior ◽  
Lipid Droplets ◽  
Lipid Domains ◽  
Intact Cells ◽  
Contact Sites ◽  
Dependent Behavior ◽  
Membrane Bound ◽  
Intracellular Vesicles ◽  
Micrometer Scale ◽  
Hypotonic Swelling

The endoplasmic reticulum (ER) is the site of synthesis of secretory and membrane proteins and contacts every organelle of the cell, exchanging lipids and metabolites in a highly regulated manner. How the ER spatially segregates its numerous and diverse functions, including positioning nanoscopic contact sites with other organelles, is unclear. We demonstrate that hypotonic swelling of cells converts the ER and other membrane-bound organelles into micrometer-scale large intracellular vesicles (LICVs) that retain luminal protein content and maintain contact sites with each other through localized organelle tethers. Upon cooling, ER-derived LICVs phase-partition into microscopic domains having different lipid-ordering characteristics, which is reversible upon warming. Ordered ER lipid domains mark contact sites with ER and mitochondria, lipid droplets, endosomes, or plasma membrane, whereas disordered ER lipid domains mark contact sites with lysosomes or peroxisomes. Tethering proteins concentrate at ER–organelle contact sites, allowing time-dependent behavior of lipids and proteins to be studied at these sites. These findings demonstrate that LICVs provide a useful model system for studying the phase behavior and interactive properties of organelles in intact cells.

scholarly journals Peroxisomal Membrane Contact Sites in Yeasts

2021 ◽  
Vol 9 ◽  
Author(s):  
Amit S. Joshi
Keyword(s):  
Lipid Droplets ◽  
Peroxisome Biogenesis ◽  
Functional Importance ◽  
Peroxisomal Membrane ◽  
Membrane Contact Sites ◽  
Contact Sites ◽  
Recent Developments ◽  
Membrane Contacts ◽  
Membrane Bound ◽  
Membrane Contact

Peroxisomes are ubiquitous, single membrane-bound organelles that play a crucial role in lipid metabolism and human health. While peroxisome number is maintained by the division of existing peroxisomes, nascent peroxisomes can be generated from the endoplasmic reticulum (ER) membrane in yeasts. During formation and proliferation, peroxisomes maintain membrane contacts with the ER. In addition to the ER, contacts between peroxisomes and other organelles such as lipid droplets, mitochondria, vacuole, and plasma membrane have been reported. These membrane contact sites (MCS) are dynamic and important for cellular function. This review focuses on the recent developments in peroxisome biogenesis and the functional importance of peroxisomal MCS in yeasts.

scholarly journals Spastin tethers lipid droplets to peroxisomes and directs fatty acid trafficking through ESCRT-III

2019 ◽  
Vol 218 (8) ◽  
pp. 2583-2599 ◽  
Author(s):  
Chi-Lun Chang ◽  
Aubrey V. Weigel ◽  
Maria S. Ioannou ◽  
H. Amalia Pasolli ◽  
C. Shan Xu ◽  
...  
Keyword(s):  
Lipid Peroxidation ◽  
Fatty Acid ◽  
Hereditary Spastic Paraplegia ◽  
Lipid Droplets ◽  
Spastic Paraplegia ◽  
Dual Roles ◽  
Aaa Atpase ◽  
Contact Sites ◽  
Membrane Morphology ◽  
Membrane Bound

Lipid droplets (LDs) are neutral lipid storage organelles that transfer lipids to various organelles including peroxisomes. Here, we show that the hereditary spastic paraplegia protein M1 Spastin, a membrane-bound AAA ATPase found on LDs, coordinates fatty acid (FA) trafficking from LDs to peroxisomes through two interrelated mechanisms. First, M1 Spastin forms a tethering complex with peroxisomal ABCD1 to promote LD–peroxisome contact formation. Second, M1 Spastin recruits the membrane-shaping ESCRT-III proteins IST1 and CHMP1B to LDs via its MIT domain to facilitate LD-to-peroxisome FA trafficking, possibly through IST1- and CHMP1B-dependent modifications in LD membrane morphology. Furthermore, LD-to-peroxisome FA trafficking mediated by M1 Spastin is required to relieve LDs of lipid peroxidation. M1 Spastin’s dual roles in tethering LDs to peroxisomes and in recruiting ESCRT-III components to LD–peroxisome contact sites for FA trafficking may underlie the pathogenesis of diseases associated with defective FA metabolism in LDs and peroxisomes.

scholarly journals Spastin tethers lipid droplets to peroxisomes and directs fatty acid trafficking through ESCRT-III

2019 ◽  
Author(s):  
Chi-Lun Chang ◽  
Aubrey V. Weigel ◽  
Maria S. Ioannou ◽  
H. Amalia Pasolli ◽  
C. Shan Xu ◽  
...  
Keyword(s):  
Lipid Peroxidation ◽  
Fatty Acid ◽  
Hereditary Spastic Paraplegia ◽  
Lipid Droplets ◽  
Spastic Paraplegia ◽  
Dual Roles ◽  
Aaa Atpase ◽  
Contact Sites ◽  
Membrane Morphology ◽  
Membrane Bound

AbstractLipid droplets (LDs) are neutral lipid storage organelles that transfer lipids to various organelles including peroxisomes. Here, we show that the hereditary spastic paraplegia protein M1 Spastin, a membrane-bound AAA ATPase found on LDs, coordinates fatty acid (FA) trafficking from LDs to peroxisomes through two inter-related mechanisms. First, M1 Spastin forms a tethering complex with peroxisomal ABCD1 to promote LD-peroxisome contact formation. Second, M1 Spastin recruits the membrane-shaping ESCRT-III proteins IST1 and CHMP1B to LDs via its MIT domain to facilitate LD-to-peroxisome FA trafficking, possibly through IST1 and CHMP1B modifying LD membrane morphology. Furthermore, M1 Spastin, IST1 and CHMP1B are all required to relieve LDs of lipid peroxidation. M1 Spastin’s dual roles in tethering LDs to peroxisomes and in recruiting ESCRT-components to LD-peroxisome contact sites for FA trafficking may help explain the pathogenesis of diseases associated with defective FA metabolism in LDs and peroxisomes.

scholarly journals ER membranes exhibit phase behavior at sites of organelle contact

2019 ◽  
Author(s):  
Christopher King ◽  
Prabuddha Sengupta ◽  
Arnold Seo ◽  
Jennifer Lippincott-Schwartz
Keyword(s):  
Endoplasmic Reticulum ◽  
Plasma Membrane ◽  
Phase Behavior ◽  
Protein Content ◽  
Cell Swelling ◽  
Lipid Domains ◽  
Contact Sites ◽  
Liquid Ordered ◽  
Er Membrane ◽  
In Cells

The plasma membrane of cells exhibits phase behavior that allows transient concentration of specific proteins and lipids, giving rise to functionally dynamic and diverse nanoscopic domains. This phase behavior is observable in giant plasma membrane-derived vesicles, in which microscopically visible, liquid-ordered (Lo) and liquid-disordered (Ld) lipid domains form upon a shift to low temperatures. The extent such phase behavior exists in the membrane of the endoplasmic reticulum (ER) of cells remains unclear. To explore the phase behavior of the ER membrane in cells, we used hypotonic cell swelling to generate Large Intra-Cellular Vesicles (LICVs) from the ER in cells. ER LICVs retained their lumenal protein content, could be retubulated into an ER network, and maintained stable inter-organelle contacts, where protein tethers are concentrated at these contacts. Notably, upon temperature reduction, ER LICVs underwent reversible phase separation into microscopically-visible Lo and Ld lipid domains. The Lo lipid domains marked ER contact sites with other organelles. These findings demonstrate that LICVs provide an important model system for studying the biophysical properties of intracellular organelles in cells.Significance StatementPrior work has demonstrated that the plasma membrane can phase separate into microscopically visible Lo and Ld domains with distinct lipid and protein content. However, such behavior on the ER membrane has not been experimentally observed, even though the ER contacts every organelle of the cell, exchanging lipids and metabolites in a highly regulated manner at these contacts. We find here that hypotonic treatment generates Large Intra-Cellular Vesicles from the endoplasmic reticulum and other membrane-bound organelles in cells, enabling the study of phase behavior on the ER membrane. We show that ER membranes can be reversibly phase separated into microscopically-observable, Lo and Ld domains. ER LICVs also maintained stable inter-organelle contact sites in cells, with organelle tethers concentrated at these contacts.

scholarly journals Seipin is required for converting nascent to mature lipid droplets

eLife ◽  
2016 ◽  
Vol 5 ◽  
Author(s):  
Huajin Wang ◽  
Michel Becuwe ◽  
Benjamin E Housden ◽  
Chandramohan Chitraju ◽  
Ashley J Porras ◽  
...  
Keyword(s):  
Lipid Droplets ◽  
Lipid Synthesis ◽  
Human Cells ◽  
Lipid Transfer ◽  
Cellular Lipid ◽  
Contact Sites ◽  
Discrete Step

How proteins control the biogenesis of cellular lipid droplets (LDs) is poorly understood. Using Drosophila and human cells, we show here that seipin, an ER protein implicated in LD biology, mediates a discrete step in LD formation—the conversion of small, nascent LDs to larger, mature LDs. Seipin forms discrete and dynamic foci in the ER that interact with nascent LDs to enable their growth. In the absence of seipin, numerous small, nascent LDs accumulate near the ER and most often fail to grow. Those that do grow prematurely acquire lipid synthesis enzymes and undergo expansion, eventually leading to the giant LDs characteristic of seipin deficiency. Our studies identify a discrete step of LD formation, namely the conversion of nascent LDs to mature LDs, and define a molecular role for seipin in this process, most likely by acting at ER-LD contact sites to enable lipid transfer to nascent LDs.

Nonlinear Lock-In Infrared Microscopy: A Complementary Investigation Technique for the Analysis of Functional Electroceramic Components

2015 ◽  
Vol 21 (5) ◽  
pp. 1145-1152 ◽  
Author(s):  
Michael Hofstätter ◽  
Nadine Raidl ◽  
Bernhard Sartory ◽  
Peter Supancic
Keyword(s):  
Electrical Properties ◽  
Electron Backscatter Diffraction ◽  
Barrier Properties ◽  
Infrared Microscopy ◽  
Polar Crystal ◽  
Dependent Behavior ◽  
Electroceramic Material ◽  
Lock In ◽  
Backscatter Diffraction ◽  
Micrometer Scale

AbstractUsing lock-in infrared microscopy as a tool for current detection on the micrometer scale in AC-driven specimens in combination with iterative grinding procedure allows preparation of current dominating microstructure regions on well-polished surfaces. This technique is applied successfully on varistor components based on specially doped ZnO-based varistor ceramics. This peculiar electroceramic material exhibits exceptional high nonlinear current–voltage (I-V) characteristics, described by a power law according I~Vα, caused by double Schottky barriers at the grain boundaries. As a novelty the thermographic response is used to evaluate local electrical properties, namely the nonlinearity coefficient α, on basis of higher order harmonics with respect to the basic electrical driving AC-frequency.To correlate the observed electrical properties to the microstructure, the polar crystal orientation of the relevant ZnO grains is determined by combining electron backscatter diffraction and orientation-dependent patterns as a result of a chemical etching procedure. These findings support a modified new model for describing the grain boundary controlled current flow in a varistor microstructure including orientation-dependent barrier properties. Hence, the experimentally observed current direction-dependent behavior can be described consistently.

scholarly journals Nanogold as a Specific Marker for Electron Cryotomography

2009 ◽  
Vol 15 (3) ◽  
pp. 183-188 ◽  
Author(s):  
Yongning He ◽  
Grant J. Jensen ◽  
Pamela J. Bjorkman
Keyword(s):  
Lipid Bilayers ◽  
Outer Surface ◽  
Three Dimensional ◽  
Fc Receptor ◽  
Specific Marker ◽  
Neonatal Fc Receptor ◽  
Membrane Bound ◽  
Nanogold Particles ◽  
Intracellular Vesicles ◽  
Electron Cryotomography

AbstractWhile electron cryotomography (ECT) provides “molecular” resolution, three-dimensional images of unique biological specimens, sample crowdedness, and/or resolution limitations can make it difficult to identify specific macromolecular components. Here we used a 1.4 nm Nanogold® cluster specifically attached to the Fc fragment of IgG to monitor its interaction with the neonatal Fc receptor (FcRn), a membrane-bound receptor that transports IgG across cells in acidic intracellular vesicles. ECT was used to image complexes formed by Nanogold-labeled Fc bound to FcRn attached to the outer surface of synthetic liposomes. In the resulting three-dimensional reconstructions, 1.4 nm Nanogold particles were distributed predominantly along the interfaces where 2:1 FcRn-Fc complexes bridged adjacent lipid bilayers. These results demonstrate that the 1.4 nm Nanogold cluster is visible in tomograms of typically thick samples (∼250 nm) recorded with defocuses appropriate for large macromolecules and is thus an effective marker.

scholarly journals A Mutant of Paracoccus denitrificans with Disrupted Genes Coding for Cytochrome c550 and Pseudoazurin Establishes These Two Proteins as the In Vivo Electron Donors to Cytochrome cd1 Nitrite Reductase

2003 ◽  
Vol 185 (21) ◽  
pp. 6308-6315 ◽  
Author(s):  
Isobel V. Pearson ◽  
M. Dudley Page ◽  
Rob J. M. van Spanning ◽  
Stuart J. Ferguson
Keyword(s):  
Nitrite Reductase ◽  
Paracoccus Denitrificans ◽  
Wild Type ◽  
Anaerobic Conditions ◽  
Electron Mediator ◽  
Intact Cells ◽  
Cytochrome Cd1 ◽  
Membrane Bound ◽  
Electron Carriers

ABSTRACT In Paracoccus denitrificans, electrons pass from the membrane-bound cytochrome bc 1 complex to the periplasmic nitrite reductase, cytochrome cd 1. The periplasmic protein cytochrome c 550 has often been implicated in this electron transfer, but its absence, as a consequence of mutation, has previously been shown to result in almost no attenuation in the ability of the nitrite reductase to function in intact cells. Here, the hypothesis that cytochrome c 550 and pseudoazurin are alternative electron carriers from the cytochrome bc 1 complex to the nitrite reductase was tested by construction of mutants of P. denitrificans that are deficient in either pseudoazurin or both pseudoazurin and cytochrome c 550. The latter organism, but not the former (which is almost indistinguishable in this respect from the wild type), grows poorly under anaerobic conditions with nitrate as an added electron acceptor and accumulates nitrite in the medium. Growth under aerobic conditions with either succinate or methanol as the carbon source is not significantly affected in mutants lacking either pseudoazurin or cytochrome c 550 or both these proteins. We concluded that pseudoazurin and cytochrome c 550 are the alternative electron mediator proteins between the cytochrome bc 1 complex and the cytochrome cd 1-type nitrite reductase. We also concluded that expression of pseudoazurin is mainly controlled by the transcriptional activator FnrP.

scholarly journals Upregulated microRNA-106a Promotes Porcine Preadipocyte Proliferation and Differentiation by Targeting Different Genes

Genes ◽  
2019 ◽  
Vol 10 (10) ◽  
pp. 805 ◽  
Author(s):  
Kuilong Huang ◽  
Xin’e Shi ◽  
Jie Wang ◽  
Ying Yao ◽  
Ying Peng ◽  
...  
Keyword(s):  
Cell Proliferation ◽  
Lipid Droplets ◽  
Luciferase Reporter ◽  
Cell Cycles ◽  
Lipogenic Genes ◽  
Proliferation And Differentiation ◽  
Luciferase Reporter Vector ◽  
Membrane Bound ◽  
Porcine Adipocytes ◽  
Oil Red O

Adipose tissue is one of the main organs for the energy storage and supply of organisms. Adipose deposition and metabolism are controlled by a cascade of transcription factors and epigenetic regulatory mechanisms. Previous studies have also shown that miR-106a plays a considerable role in the development of organisms. The regulatory mechanism of miR-106a on porcine preadipocytes is still not clear. In this study, preadipocytes were isolated from the neck subcutaneous deposits of 3–5-day old Chinese native Guanzhong black pigs using 5-ethynyl-20-deoxyuridine (EdU) staining and a CCK-8 assay to detect the number of proliferous cells and real-time qPCR (RT-qPCR) and western blot analysis to detect gene expression, as well as Oil Red O and BODIPY staining dye lipid droplets and flow cytometry (FCM) to detect cell cycles. We also used the double luciferase method to detect the relative luciferase activities. Upregulated miR-106a increased the number of proliferous cells and enhanced the expression of cell proliferation-related genes in porcine adipocytes. The double luciferase reporter vector confirmed that p21 was a target gene of miR-106a in the cell proliferation phase. miR-106a upregulation increased the number of lipid droplets and the expression of lipogenic genes and directly targeted BMP and activin membrane-bound inhibitor (BAMBI) in the process of differentiation. Our results indicated that miR-106a promotes porcine preadipocyte proliferation and differentiation by targeting p21 and BAMBI.

Effect of membrane perturbing treatments on the membrane-bound peptidases ofStreptococcus cremorisHP

1979 ◽  
Vol 46 (3) ◽  
pp. 473-484 ◽  
Author(s):  
Fred A. Exterkate
Keyword(s):  
Intact Cells ◽  
Irreversible Inhibition ◽  
Alkaline Conditions ◽  
Membrane Bound ◽  
Functional Peptide ◽  
Solvent Molecules ◽  
And Storage ◽  
No Activation ◽  
Proline Iminopeptidase

SummaryThe effects of solubilization, treatment with organic solvents and storage under alkaline conditions on membrane-associated peptidases of intact cells ofStreptococcus cremorisHP were studied. Differences in the response of the peptidase activities towards these membrane perturbing treatments were observed. Pyrrolidonecarboxylylpeptidase (PCP) and an endopeptidase (P50) showed 50% irreversible inhibition at the same concentration of each solvent tested. An amino- and proline iminopeptidase activity and the endopeptidase P37were in this respect much more sensitive to the action of the solvents. Within a homologous series of n-alkanols irreversible inhibition of PCP showed a dependence on the hydrophobicity of the solvent molecules. Only P37activity was increased considerably upon solubilization of the enzyme. Similar levels of activation were found upon treatment of cells with 3% (v/v) n-butanol at 25 °C or storage at 30 °C at an alkaline pH. Optimal activity of P50during n-butanol treatment was at 25 °C using a concentration of 5% (v/v), but no activation was observed upon solubilization. The results are discussed in terms of enzyme–lipid interaction and accessibility of the enzymes in situ. It is concluded that the enzymes apparently occupy different positions within the membrane although they may together constitute a functional peptide-hydrolysing unit.

Sign in / Sign up

Export Citation Format

Share Document