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Cholesterol transport through the peroxisome-ER membrane contacts tethered by PI（4,5）P2 and extended synaptotagmins

Science China(Life Sciences) 2019年第9期62卷 1117-1135页

作者： Jian Xiao Jie Luo Ao Hu Ting Xiao Meixin Li Zekai Kong Luyi Jiang Zimu Zhou Yacheng Liao Chang Xie Beibei Chu Honghua Miao Boliang Li Xiongjie Shi Bao-Liang Song College of Life Sciences the Institute for Advanced StudiesWuhan UniversityWuhan 430072China State Key Laboratory of Molecular Biology Institute of Biochemistry and Cell BiologyShanghai Institutes for Biological SciencesChinese Academy of SciencesShanghai 200031China Shenzhen Institute of Wuhan University Shenzhen 518057China

Most mammalian cells take up cholesterol from low-density lipoproteins(LDLs) via receptor-mediated endocytosis.After reaching lysosomes,LDL-derived cholesterol continues to transport to downstream organelles including the ER for specific structural and functional needs.Peroxisomes are recently found to receive cholesterol from lysosomes through lysosomeperoxisome membrane contacts.However,whether and how cholesterol is conveyed from peroxisomes to the ER remain unknown.Here,by combining high-resolution microscopic analyses and in vitro reconstitution of highly purified organelles or artificial liposomes,we demonstrate that peroxisomes form membrane contacts with the ER through the interaction between peroxisomal PI(4,5)P2 and ER-resident extended synaptotagmin-1,2 and 3(E-Syts).Depletion of peroxisomal PI(4,5)P2 or ESyts markedly decreases peroxisome-ER membrane contacts and induces cholesterol accumulation in lysosomes.Furthermore,we show that cholesterol is delivered from 3H-labeled peroxisomes or PI(4,5)P2-containing liposomes to the ER in vitro,and that the presence of peroxisomes augments cholesterol transfer from lysosomes to the ER.Together,our study reveals a new cholesterol transport pathway along the lysosome-peroxisome-ER membrane contacts in the cell.

关键词： Peroxisome ER membrane contacts E-Syts PI(4,5)P2

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Comparison of CIM~ C4-HLD monolithic column with Sartobind phenyl membrane column for pIDKE2 purification

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色谱 2017年第10期35卷 1028-1036页

作者： Miladys LIMONTA Lourdes ZUMALACRREGUI Urska VIDIC Nika LENDERO KRAJNC Center for Genetic Engineering and Biotechnology Technical University of Havana"José Antonio Echeverría" BIA Separations

The main component of the Center for Genetic Engineering and Biotechnology(CIGB)candidate vaccine against Hepatitis C virus(HCV)is the pIDKE2 plasmid.The current designed downstream process for the production of pIDKE2 fulfils all regulatory requirements and renders the required quantities of pharmaceuticalgrade plasmid DNA(pDNA)with 95%purity.The advantages of this procedure include high plasmid purity and the elimination of undesirable additives,such as toxic organic extractants and animal-derived enzymes.However,yields and consequently the productivity of the process are low.Previous work demonstrated that the most critical step of the process is the reverse phase chromatography,where conventional porous particle resins are used.Therefore,to increase the process productivity,alternative technologies such as membranes and chromatographic monoliths were tested as alternative options for this critical step.Here,a comparison between the behaviors of CIM~ C4-HLD and Sartobind phenyl matrices was performed.To obtain higher productivities and purities,the dynamic binding capacities and selectivities were evaluated.The results showed that both matrices had a similar capacity for pIDKE2 plasmid,but the separation of pDNA isoforms using CIM~ technology was much better than that with Sartobind.Additionally,the optimal conditions for loading plasmid DNA on a CIMC4-HLD 800-mL monolithic column in a real production process were determined.These optimizations will allow production levels to satisfy the high plasmid consumption demanded by clinical trials.

关键词： membrane monolith downstream processing plasmid DNA (pDNA) breakthrough supercoiled (SC) plasmid open circular (OC) plasmid

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Red blood cell membrane-camouflaged nanoparticles: a novel drug delivery system for antitumor application

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Acta Pharmaceutica Sinica B 2019年第4期9卷 675-689页

作者： Qing Xia Yongtai Zhang Zhe Li Xuefeng Hou Nianping Feng Department of Pharmaceutical Sciences Shanghai University of Traditional Chinese Medicine

Erythrocytes(red blood cells, RBCs) are the most abundant circulating cells in the blood and have been widely used in drug delivery systems(DDS) because of their features of biocompatibility,biodegradability, and long circulating half-life. Accordingly, a 'camouflage' comprised of erythrocyte membranes renders nanoparticles as a platform that combines the advantages of native erythrocyte membranes with those of nanomaterials. Following injection into the blood of animal models, the coated nanoparticles imitate RBCs and interact with the surroundings to achieve long-term circulation. In this review, the biomimetic platform of erythrocyte membrane-coated nano-cores is described with regard to various aspects, with particular focus placed on the coating mechanism, preparation methods, verification methods, and the latest anti-tumor applications. Finally, further functional modifications of the erythrocyte membranes and attempts to fuse the surface properties of multiple cell membranes are discussed,providing a foundation to stimulate extensive research into multifunctional nano-biomimetic systems.

关键词： Red blood cells membrane Nanoparticles Drug delivery Antitumor Biomimetic nanoparticles

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Cholesterol-tuned liposomal membrane rigidity directs tumor penetration and anti-tumor effect

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Acta Pharmaceutica Sinica B 2019年第4期9卷 858-870页

作者： Hangyi Wu Miaorong Yu Yunqiu Miao Shufang He Zhuo Dai Wenyi Song Yuan Liu Sha Song Ejaj Ahmad Dongkai Wang Yong Gan Department of Pharmaceutics Shenyang Pharmaceutical UniversityShenyang 117004China Shanghai Institute of Materia Medica Chinese Academy of SciencesShanghai 201203China

Recently, liposomes have been widely used in cancer therapeutics, but their anti-tumor effects are suboptimal due to limited tumor penetration. To solve this problem, researchers have made significant efforts to optimize liposomal diameters and potentials, but little attention has been paid to liposomal membrane rigidity. Herein, we sought to demonstrate the effects of cholesterol-tuned liposomal membrane rigidity on tumor penetration and anti-tumor effects. In this study, liposomes composed of hydrogenated soybean phospholipids(HSPC), 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000](DSPE-PEG2000) and different concentrations of cholesterol were prepared. It was revealed that liposomal membrane rigidity decreased with the addition of cholesterol. Moderate cholesterol content conferred excellent diffusivity to liposomes in simulated diffusion medium, while excessive cholesterol limited the diffusion process. We concluded that the differences of the diffusion rates likely stemmed from the alterations in liposomal membrane rigidity, with moderate rigidity leading to improved diffusion. Next, the in vitro tumor penetration and the in vivo anti-tumor effects were analyzed. The results showed that liposomes with moderate rigidity gained excellent tumor penetration and enhanced anti-tumor effects. These findings illustrate a feasible and effective way to improve tumor penetration and therapeutic efficacy of liposomes by changing the cholesterol content, and highlight the importance of liposomal membrane rigidity.

关键词： Cholesterol Liposomal membrane rigidity Tumor penetration Anti-tumor effects

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Proteomics analysis of Trichoplusia ni midgut epithelial cell brush border membrane vesicles

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Insect Science 2019年第3期26卷 424-440页

作者： Muhammad Afzal Javed Cathy Coutu David A. Theilmann Martin A. Erlandson Dwayne D. Hegedus Saskatoon Research and Development Centre Agriculture and Agri-Food Canada Saskatoon Saskatchewan Canada Summerland Research and Development Centre Agriculture and Agri-Food Canada Summerland British Columbia Canada Department of Food & Bio-Product Sciences University of Saskatchewan Saskatoon Saskatchewan Canada

The insect midgut epithelium is composed of columnar, goblet, and regenerative cells. Columnar epithelial cells are the most abundant and have membrane protrusions that form the brush border membrane (BBM) on their apical side. These increase surface area available for the transport of nutrients, but also provide opportunities for interaction with xenobiotics such as pathogens, toxins and host plant allelochemicals. Recent improvements in proteomic and bioinfbrmatics tools provided an opportunity to determine the proteome of the T. ni BBM in unprecedented detail. This study reports the identification of proteins from BBM vesicles (BBMVs) using single dimension polyacrylamide gel elec? trophoresis coupled with multi-dimensional protein identification technology. More than 3000 proteins were associated with the BBMV of which 697 were predicted to possess either a signal peptide, at least one transmembrane domain or a GPI-anchor signal. Of these, bioinfbrmatics analysis and manual curation predicted that 185 may be associated with the BBMV or epithelial cell plasma membrane. These are discussed with respect to their predicted functions, namely digestion, nutrient uptake, cell signaling, development, cell-cell interactions, and other functions. We believe this to be the most detailed proteomic analysis of the lepidopteran midgut epithelium membrane to date, which will provide information to better understand the biochemical, physiological and pathological processes taking place in the larval midgut.

关键词： brush border membrane vesicles midgut proteomic analysis Trichoplusiani

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Effect of Pinellia ternata Lectin on membrane Currents of Mouse Motor Nerve Terminals

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Science China Chemistry 1994年第4期37卷 448-453页

作者：施玉樑徐幼芬张辉 Shanghai Institute of Physiology Academia Sinica Shanghai 200031 PRC

Pinellia ternata lectin (PTL) extracted from the fresh juice of rhizome of pinellia ternata used as a traditional Chinese medicine facilitated the quantal release of acetylcholine (ACh) in the mouse motor nerve terminals and formed cation channels in artificial lipid bilayer. Here we report the action of PTL on presynaptic membrane currents of motor nerve terminals.The experiments were performed on the intercostal nerve triangularis sterni muscle preparations. By means of the perineurial recording, the effects of PTL on the sodium current in the preterminal part , three potassium currents and two calcium currents generated from the nerve terminals were investigated. The results show that PTL increases voltage-dependent fast Ca2+ current (ICa,f), Na+ current (INa) and Ca2+-acti-vated K+ current (IK,Ca) without action on either the voltage-dependent fast K+ current (IK,f) or the slow K+ current (IK,S). These effects are irreversible, but can be reversed by mannan, the specific binding sugar for PTL.The

关键词： Pinellia ternata lectin motor nerve terminal peYineurial recording membrane currents.

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Crystal structure of kindlin-2 PH domain reveals a conformational transition for its membrane anchoring and regulation of integrin activation

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Protein & Cell 2012年第6期3卷 434-440页

作者： Yan Liu Yun Zhu Sheng Ye Rongguang Zhang National Laboratory of Biophysics Institute of BiophysicsChinese Academy of SciencesBeijing 100101China

Kindlin-2 belongs to a subfamily of FERM domain con-taining proteins,which plays key roles in activating integrin transmembrane receptors and mediating cell adhesion.Compared to conventional FERM domains,kindlin-2 FERM contains an inserted pleckstrin homology(PH)domain that specifically binds to phosphatidy-linositol(3,4,5)trisphosphate(PIP3)and regulates the kindlin-2 function.We have determined the crystal struc-ture of kindlin-2 PH domain at 1.9Åresolution,which reveals a conserved PH domain fold with a highly charged and open binding pocket for PIP3 head group.Structural comparison with a previously reported solution structure of kindlin-2 PH domain bound to PIP3 head group reveals that upon PIP3 insertion,there is a significant conformational change of both the highly positively charged loop at the entry of the PIP3 binding pocket and the entireβbarrel of the PH domain.We propose that such“induced-fit”type change is crucial for the tight binding of PIP3 to anchor kindlin-2 onto the membrane surface,thereby promoting its binding to integrins.Our results provide important structural insight into kindlin-2-mediated membrane anchoring and integrin activation.

关键词： kindlin-2 integrin PH domain crystal structure cell adhesion membrane

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Venovenous extra-corporeal membrane oxygenation for severe acute respiratory distress syndrome: a matched cohort study

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Chinese Medical Journal 2019年第18期132卷 2192-2198页

作者： Song-Qiao Liu Ying-Zi Huang Chun Pan Lan-Qi Guo Xiao-Ting Wang Wen-Kui Yu Yun-Fu Wu Jie Yan Hong-Sheng Zhao Ling Liu Feng-Mei Guo Jing-Yuan Xu Yi Yang Hai-Bo Qiu Department of Critical Care Medicine Zhongda Hospital School of Medicine Southeast University Nanjing Jiangsu 210009 China Department of Critical Care Medicine Peking Union Medical College Hospital Peking Union Medical College Chinese Academy of Medical Science Beijing 100010 China Department of General Surgery Jinling Hospital Medical School of Nanjing University Nanjing Jiangsu 210002 China Department of Critical Care Medicine Suzhou Hospital Affiliated to Nanjing Medical University Suzhou Jiangsu 215001 China Department of Critical Care Medicine Wuxi People’s Hospital Nanjing Medical University Wuxi Jiangsu 214023 China Department of Critical Care Medicine Hospital of Nantong University Nantong Jiangsu 226001 China

Background:Although the use of extra-corporeal membrane oxygenation (ECMO) has been rapidly increasing,the benefit of ECMO in patients with acute respiratory distress syndrome (ARDS) remains unclear.Our objective was to investigate the effect of venovenous ECMO (VV-ECMO) on adult patients with severe ARDS.Methods:We conducted a multi-center,retrospective,cohort study in the intensive care units (ICUs) of six teaching hospitals between January 2013 and December 2018.Patients with severe ARDS who received W-ECMO support were included.The detailed demographic data and physiologic data were used to match ARDS patients without ECMO.The primary endpoint was the 28-day mortality.Results:Ninety-nine patients with severe ARDS supported by VV-ECMO and 72 patients without ECMO were included in this study.The acute physiology and chronic health evaluation Ⅱ score was 23.1 ± 6.3 in the ECMO group and 24.8 ± 8.5 in the control group (P =0.1195).The sequential organ failure assessment score was 12.8 ± 3.4 in the ECMO group and 13.7 ± 3.5 in the control group (P =0.0848).The 28-day mortality of patients with ECMO support was 39.4%,and that of the control group was 55.6%.The survival analysis curve showed that the 28-day mortality in the ECMO group was significantly lower than that in the control group (P =0.0097).Multivariate Cox regression analysis showed that the independent predictors of the 28-day mortality were the requirement of vasopressors before ECMO (hazard ratio [HR]:1.006;95% confidence interval [CI]:1.001-1.013;P =0.030) and duration of mechanical ventilation before ECMO (HR:3.299;95% CI:1.264-8.609;P =0.034).Conclusions:This study showed that ECMO improved the survival of patients with severe ARDS.The duration of mechanical ventilation and the requirement of vasopressors before ECMO might be associated with an increased risk of death.

关键词： Acute respiratory distress syndrome Extra-corporeal membrane oxygenation Mortality Multi-center Mechanical ventilation

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Study on screening potential allergenic proteins from infant milk powders based on human mast cell membrane chromatography and histamine release assays

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Journal of Pharmaceutical Analysis 2019年第1期9卷 55-61页

作者： Ping Zhang Yingdi Shi Xiaoshuang He Wei Sun Yanni Lv Xiaofang Hou School of Pharmacy Xi'an Jiaotong University Linyi Institute for Food and Drug Control Shaanxi Engineering Research Center of Cardiovascular Drugs Screening & Analysis

Cow's milk allergy is mainly observed in infants and young children. Most allergic reactions affect the skin, followed by the gastrointestinal and respiratory systems. Conventional diagnosis is based on positive allergy studies and evaluation of parameters including IgE and IgG1 levels, acute allergic skin response and anaphylactic shock reactions. We developed a cell membrane chromatographic(CMC)method based on human mast cells(HMC-1) for screening potential allergens in infant formula milk powders(IFMP). HMC-1 cell membranes were extracted and mixed with silica to prepare cell membrane chromatography columns(10 mm ? 2 mm i.d., 5 mm). Under the conditions of 0.2 mL/min flow rate and214 nm detection wavelength, human breast milk showed no retention. However, IFMP showed clear retention. The retained fractions were collected and analyzed through matrix-assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF-MS). Four major milk proteins, i.e., α-casein, β-casein, α-lactalbumin, and β-lactoglobulin A, were identified. Furthermore, these proteins and β-lactoglobulin B showed clear retention on HMC-1/CMC columns. To test the degranulation effects of the five proteins, histamine and β-hexosaminidase release assays were carried out. All five proteins induced HMC-1 cells to release histamine and β-hexosaminidase. Also, we established a reversed phase liquid chromatographic(RPLC) method for the determination of the five proteins in IFMP and the results showed that 90% proteins in IFMP were α-casein and β-casein. We concluded that cow's milk proteins may be potential allergens and caseins cause more β-casein allergic risk than other proteins. This conclusion was consistent with other studies.

关键词： Allergenic proteins Cell membrane chromatography Milk powders

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Topology-empowered membrane devices for terahertz photonics

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Advanced Photonics 2022年第4期4卷 29-37页

作者： Quanlong Yang Dongyang Wang Sergey Kruk Mingkai Liu Ivan Kravchenko Jiaguang Han Yuri Kivshar Ilya Shadrivov Australian National University Nonlinear Physics CentreResearch School of PhysicsCanberra ActonAustralia Central South University School of Physics and ElectronicsChangshaChina The Hong Kong University of Science and Technology Department of PhysicsHong KongChina Center for Nanophase Materials Sciences Oak Ridge National LaboratoryOak RidgeTennesseeUnited States Tianjin University Center for Terahertz Waves and College of Precision Instrument and Optoelectronics EngineeringTianjinChina

Control of terahertz waves offers a profound platform for next-generation sensing,imaging,and information communications.However,all conventional terahertz components and systems suffer from bulky design,sensitivity to imperfections,and transmission loss.We propose and experimentally demonstrate onchip integration and miniaturization of topological devices,which may address many existing drawbacks of the terahertz technology.We design and fabricate topological devices based on valley-Hall photonic structures that can be employed for various integrated components of on-chip terahertz systems.We demonstrate valleylocked asymmetric energy flow and mode conversion with topological waveguide,multiport couplers,wave division,and whispering gallery mode resonators.Our devices are based on topological membrane metasurfaces,which are of great importance for developing on-chip photonics and bring many features into terahertz technology.

关键词： topological devices terahertz photonics membrane on-chip

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