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Evaluation of the reducing potential of PSMA-containing endosomes by FRET imaging

Cancer Drug Resistance 2021年第1期4卷 223-232页

作者： Chelvam Venkatesh Jiayin Shen Karson S.Putt Philip S.Low Discipline of Chemistry Indian Institute of TechnologyMadhya PradeshIndore 453552India Discipline of Bioscience and Biomedical Engineering Indian Institute of TechnologyMadhya PradeshIndore 453552India Department of Chemistry Purdue UniversityWest LafayetteIN 47907USA Institute for Drug Discovery Purdue UniversityWest LafayetteIN 47907USA

Aim:Ligand-targeted therapeutics are experiencing increasing use for treatment of human diseases due to their ability to concentrate a desired drug at a pathologic site while reducing accumulation in healthy *** many ligand-targeted drug conjugates,a critical aspect of conjugate design lies in engineering release of the therapeutic payload to occur only after its internalization by targeted *** disulfide bond reduction is frequently exploited to ensure intracellular drug release,an understanding of the redox properties of endocytic compartments can be critical to ligand-targeted drug *** the redox properties of folate receptor trafficking endosomes have been previously reported,little is known about the trafficking of prostate-specific membrane antigen(PSMA),a receptor that is experiencing increasing use for drug targeting in ***:To obtain this information,we have constructed a PSMA-targeted fluorescence resonance energy transfer pair that reports on disulfide bond reduction by changing fluorescence from red to ***:We show here that this reporter exhibits rapid and selective uptake by PSMA-positive cells,and that reduction of its disulfide bond proceeds steadily but incompletely following *** fact that maximal disulfide reduction reaches only~50%,even after 24 h incubation,suggests that roughly half of the conjugates must traffic through endosomes that display no reducing ***:As the level of disulfide reduction differs between PSMA trafficked and previously published folate trafficked conjugates,it also follows that not all internalizing receptors are translocated through similar intracellular *** together,these data suggest that the efficiency of disulfide bond reduction must be independently analyzed for each receptor trafficking pathway when disulfide bond reduction is exploited for intracellular drug release.

关键词： DUPA prostate-specific membrane antigen endosomes endocytosis

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New aspects of a small GTPase RAB35 in brain development and function

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Neural Regeneration Research 2025年第7期20卷 1971-1980页

作者： Ikuko Maejima Ken Sato Laboratory of Molecular Traffic Institute for Molecular and Cellular RegulationGunma UniversityMaebashiJapan

In eukaryotic cells,organelles in the secretory,lysosomal,and endocytic pathways actively exchange biological materials with each other through intracellular membrane trafficking,which is the process of transporting the cargo of proteins,lipids,and other molecules to appropriate compartments via transport vesicles or *** processes are strictly regulated by various small GTPases such as the RAS-like in rat brain(RAB)protein family,which is the largest subfamily of the RAS *** of membrane trafficking affects tissue homeostasis and leads to a wide range of diseases,including neurological disorders and neurodegenerative ***,it is important to understand the physiological and pathological roles of RAB proteins in brain ***35,a member of the RAB family,is an evolutionarily conserved protein in metazoans.A wide range of studies using cultured mammalian cells and model organisms have revealed that RAB35 mediates various processes such as cytokinesis,endocytic recycling,actin bundling,and cell ***35 is also involved in neurite outgrowth and turnover of synaptic *** generated brain-specific Rab35 knockout mice to study the physiological roles of RAB35 in brain development and *** mice exhibited defects in anxiety-related behaviors and spatial ***,RAB35 is required for the precise positioning of pyramidal neurons during hippocampal development,and thereby for normal hippocampal *** contrast,layer formation in the cerebral cortex occurred superficially,even in the absence of RAB35,suggesting a predominant role for RAB35 in hippocampal development rather than in cerebral cortex *** studies have suggested an association between RAB35 and neurodegenerative diseases,including Parkinson's disease and Alzheimer's *** this review,we provide an overview of the current understanding of subcellular functions of *** also provide insights into the phys

关键词： endocytosis endosomes hippocampal development neurodegenerative diseases RAB35

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SARS-CoV-2 treatment effects induced by ACE2-expressing microparticles are explained by the oxidized cholesterolincreased endosomal pH of alveolar macrophages

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Cellular & Molecular Immunology 2022年第2期19卷 210-221页

作者： Zhenfeng Wang Jiadi Lv Pin Yu Yajin Qu Yabo Zhou Li Zhou Qiangqiang Zhu Shunshun Li Jiangping Song Wei Deng Ran Gao Yuying Liu Jiangning Liu Wei-Min Tong Chuan Qin Bo Huang Department of Immunology&National Key Laboratory of Medical Molecular Biology Institute of Basic Medical SciencesChinese Academy of Medical Sciences(CAMS)&Peking Union Medical CollegeBeijing 100005China NHC Key Laboratory of Human Disease Comparative Medicine Beijing Key Laboratory for Animal Models of Emerging and Remerging Infectious DiseasesInstitute of Laboratory Animal ScienceCAMS and Comparative Medicine CenterPeking Union Medical CollegeBeijingChina State Key Laboratory of Cardiovascular Disease Fuwai HospitalNational Center for Cardiovascular DiseasesCAMS and Peking Union Medical CollegeBeijingChina Department of Pathology Institute of Basic Medical SciencesCAMS and Peking Union Medical CollegeBeijingChina Department of Biochemistry&Molecular Biology Tongji Medical CollegeHuazhong University of Science&TechnologyWuhan 430030China

Exploring the cross-talk between the immune system and advanced biomaterials to treat SARS-CoV-2 infection is a promising ***,we show that ACE2-overexpressing A549 cell-derived microparticles(AO-MPs)are a potential therapeutic agent against SARS-CoV-2 *** administered AO-MPs dexterously navigate the anatomical and biological features of the lungs to enter the alveoli and are taken up by alveolar macrophages(AMs).Then,AO-MPs increase the endosomal pH but decrease the lysosomal pH in AMs,thus escorting bound SARS-CoV-2 from phago-endosomes to lysosomes for *** pH regulation is attributable to oxidized cholesterol,which is enriched in AO-MPs and translocated to endosomal membranes,thus interfering with proton pumps and impairing endosomal *** addition to promoting viral degradation,AO-MPs also inhibit the proinflammatory phenotype of AMs,leading to increased treatment efficacy in a SARS-CoV-2-infected mouse model without side *** findings highlight the potential use of AO-MPs to treat SARS-CoV-2-infected patients and showcase the feasibility of MP therapies for combatting emerging respiratory viruses in the future.

关键词： SARS-CoV-2 microparticles Alveolar macrophages endosomes Lysosomes

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The convergent point of the endocytic and autophagicpathways in leydig cells

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Cell Research 1999年第4期9卷 243-253页

作者： YI JING XUE MING TANG(Department of Biophysics and Cell Biology, Shanghai Second Medical University, Shanghai 200025, China) Department of Biophysics and Cell Biology Shanghai Second Medical University Shanghai 200025 China

INTRODUCTIONEndocytosisandalltophagocytosis(alltophagy)arethemajorpathwaysformaterialstobetransportedtolysosolnesillanimalcells,theformerbeingresponsibleforuptakeofextracellular***... 详细信息

INTRODUCTIONEndocytosisandalltophagocytosis(alltophagy)arethemajorpathwaysformaterialstobetransportedtolysosolnesillanimalcells,theformerbeingresponsibleforuptakeofextracellular***...

关键词： Autophagocytosis endocytosis endosomes lysosomes.

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Targeting γ-secretase triggers the selective enrichment of oligomeric APP-CTFs in brain extracellular vesicles from Alzheimer cell and mouse models

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Translational Neurodegeneration 2019年第1期8卷 433-449页

作者： Inger Lauritzen Anaïs Bécot Alexandre Bourgeois Raphaëlle Pardossi-Piquard Maria-Grazia Biferi Martine Barkats Fréderic Checler Institut de Pharmacologie Moléculaire et Cellulaire CNRS-UMR7275team labeled«Fondation pour la Recherche Médicale»et«Laboratoire d’excellence Distalz»Universitéde Nice-Sophia-AntipolisSophia-AntipolisFrance Institut-Myologie ParisFrance

Background:We recently demonstrated an endolysosomal accumulation of theβ-secretase-derived APP C-terminal fragment(CTF)C99 in brains of Alzheimer disease(AD)mouse ***,we showed that the treatment with theγ-secretase inhibitor(D6)led to further increased endolysosomal APP-CTF levels,but also revealed extracellular APP-CTF-associated *** here hypothesized that this latter staining could reflect extracellular vesicle(EV)-associated APP-CTFs and aimed to characterize theseγ-secretase inhibitor-induced ***:EVs were purified from cell media or mouse brains from vehicle-or D6-treated C99 or APPswedish expressing cells/mice and analyzed for APP-CTFs by *** pharmacological,immunological and genetic approaches(presenilin invalidation and C99 dimerization mutants(GXXXG))were used to characterize vesicle-containing *** APP-CTF localization was determined by ***:Purified EVs from both AD cell or mouse models were enriched in APP-CTFs as compared to EVs from control cells/***,EVs from D6-treated cells not only displayed increased C99 and C99-derived C83 levels but also higher molecular weight(HMW)APP-CTF-immunoreactivities that were hardly detectable in whole cell ***,the intracellular levels of HMW APP-CTFs were amplified by the exosomal inhibitor *** combined pharmacological,immunological and genetic approaches,we established that these HMW APP-CTFs correspond to oligomeric APP-CTFs composed of C99 and/or *** analysis showed that monomers were localized mainly to the trans-Golgi network,whereas oligomers were confined to endosomes and lysosomes,thus providing an anatomical support for the selective recovery of HMW APP-CTFs in *** D6-induced APP-CTF oligomerization and subcellular mislocalization was indeed due toγ-secretase blockade,since it similarly occurred in presenilin-deficient ***,our data proposed t

关键词： Extracellular vesicles C99 APP-CTFs Homo-and hetero-oligomerization endosomes Lysosomes trans-Golgi network SorLA γ-Secretase inhibition Presenilin knockout Alzheimer’s disease

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Upregulation of RIN3 induces endosomal dysfunction in Alzheimer’s disease

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Translational Neurodegeneration 2020年第2期9卷 290-308页

作者： Ruinan Shen Xiaobei Zhao Lu He Yongbo Ding Wei Xu Suzhen Lin Savannah Fang Wanlin Yang Kijung Sung Brian Spencer Robert A.Rissman Ming Lei Jianqing Ding Chengbiao Wu Institute of Neurology Ruijing HospitalShanghai JiaoTong University School of Medicine197 Ruijin Er Rd.Shanghai200025China Department of Neurosciences University of California San Diego School of MedicineRoom 312 MC-06249500 Gilman DriveLa JollaCA92093-0624USA Shanghai Institute of Precision Medicine Shanghai200125China Department of Neurology Zhuijiang HospitalSouthern Medical UniversityGuangzhouChina San Diego VA Health System San DiegoCAUSA

Background In Alzheimer’s Disease(AD),about one-third of the risk genes identified by GWAS encode proteins that function predominantly in the endocytic *** them,the Ras and Rab Interactor 3(RIN3)is a guanine nucleotide exchange factor(GEF)for the Rab5 small GTPase family and has been implicated to be a risk factor for both late onset AD(LOAD)and sporadic early onset AD(sEOAD).However,how RIN3 is linked to AD pathogenesis is currently *** Quantitative PCR and immunoblotting were used to measure the RIN3 expression level in mouse brain tissues and cultured basal forebrain cholinergic neuron(BFCNs).Immunostaining was used to define subcellular localization of RIN3 and to visualize endosomal changes in cultured primary BFCNs and PC12 *** flag-tagged RIN3 protein was purified from HEK293T cells and was used to define RIN3-interactomes by mass ***3-interacting partners were validated by co-immunoprecipitation,immunofluorescence and yeast two hybrid *** imaging of primary neurons was used to examine axonal transport of amyloid precursor protein(APP)andβ-secretase 1(BACE1).Immunoblotting was used to detect protein expression,processing of APP and phosphorylated forms of *** We have shown that RIN3 mRNA level was significantly increased in the hippocampus and cortex of APP/PS1 mouse *** forebrain cholinergic neurons(BFCNs)cultured from E18 APP/PS1 mouse embryos also showed increased RIN3 expression accompanied by early endosome *** addition,via its proline rich domain,RIN3 recruited BIN1(bridging integrator 1)and CD2AP(CD2 associated protein),two other AD risk factors,to early ***,overexpression of RIN3 or CD2AP promoted APP cleavage to increase its carboxyl terminal fragments(CTFs)in PC12 *** of RIN3 or the neuronal isoform of BIN1 increased phosphorylated Tau ***,upregulation of RIN3 expression promoted accumulation of APP CTFs and increased

关键词： Alzheimer's disease(AD) AD risk factors endosomes Trafficking RIN3 BIN1 CD2AP Tau

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Interplay of transport vesicles during plant-fungal pathogen interaction

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Stress Biology 2023年第1期3卷 377-386页

作者： Yakubu Saddeeq Abubakar Idris Zubair Sadiq Aarti Aarti Zonghua Wang Wenhui Zheng State Key Laboratory of Ecological Pest Control for Fujian and Taiwan Crops College of Life SciencesFujian Agriculture and Forestry UniversityFuzhouChina Key Laboratory of Integrated Pest Management for Fujian-Taiwan Crops Ministry of AgricultureCollege of Plant ProtectionFujian Agriculture and Forestry UniversityFuzhouChina Department of Biochemistry Faculty of Life SciencesAhmadu Bello UniversityZariaNigeria Fuzhou Institute of Oceanography Minjiang UniversityFuzhouChina

Vesicle trafficking is an essential cellular process upon which many physiological processes of eukaryotic cells *** is usually the‘language’of communication among the components of the endomembrane system within a cell,between cells and between a cell and its external ***,cells have the potential to internalize membrane-bound vesicles from external sources by *** constantly interact with both mutualistic and pathogenic microbes.A large part of this interaction involves the exchange of transport vesicles between the plant cells and the ***,in a pathogenic interaction,the pathogen releases vesicles containing bioactive molecules that can modulate the host immunity when absorbed by the host *** response to this attack,the host cells similarly mobilize some vesicles containing pathogenesis-related compounds to the pathogen infection site to destroy the pathogen,prevent it from penetrating the host cell or annul its *** fact,vesicle trafficking is involved in nearly all the strategies of phytopathogen attack subsequent plant immune ***,this field of plant-pathogen interaction is still at its infancy when narrowed down to plant-fungal pathogen interaction in relation to exchange of transport ***,we summarized some recent and novel findings unveiling the involvement of transport vesicles as a crosstalk in plant-fungal phytopathogen interaction,discussed their significance and identified some knowledge gaps to direct future research in the *** roles of vesicles trafficking in the development of both organisms are also established.

关键词： endosomes Extracellular vesicles Plant-pathogen interaction Phytopathogens Vesicles trafficking

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Endocytic Trafficking towards the Vacuole Plays a Key Role in the Auxin Receptor SCF^TIR-Independent Mechanism of Lateral Root Formation in A. thaliana

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Molecular Plant 2012年第6期5卷 1195-1209页

作者： Patricio Perez-Henrique Natasha V. Raikhel Lorena Norambuena Plant Molecular Biology Laboratory Department of Biology Faculty of Sciences University of Chile Santiago Chile Millenium Nucleus in Plant Cell Biotechnology Chile Center for Plant Cell Biology and Department of Botany and Plant Sciences University of California Riverside CA 92521 USA

Plants＇ developmental plasticity plays a pivotal role in responding to environmental conditions. One of the most plastic plant organs is the root system. Different environmental stimuli such as nutrients and water deficiency may induce lateral root formation to compensate for a low level of water and/or nutrients. It has been shown that the hor- mone auxin tunes lateral root development and components for its signaling pathway have been identified. Using chemi- cal biology, we discovered an Arabidopsis thaliana lateral root formation mechanism that is independent of the auxin receptor SCFTM. The bioactive compound Sortin2 increased lateral root occurrence by acting upstream from the morpho- logical marker of lateral root primordium formation, the mitotic activity. The compound did not display auxin activity. At the cellular level, Sortin2 accelerated endosomal trafficking, resulting in increased trafficking of plasma membrane recy- cling proteins to the vacuole. Sortin2 affected Late endosome/PVC/MVB trafficking and morphology. Combining Sortin2 with well-known drugs showed that endocytic trafficking of Late E/PVC/MVB towards the vacuole is pivotal for Sortin2- induced SCFTm-independent lateral root initiation. Our results revealed a distinctive role for endosomal trafficking in the promotion of lateral root formation via a process that does not rely on the auxin receptor complex SCFTM.

关键词： lateral root development endocytic trafficking Sortin2 endosomes Arabidopsis.

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Important relationships between Rab and MICAL proteins in endocytic trafficking

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World Journal of Biological Chemistry 2010年第8期1卷 254-264页

作者： Juliati Rahajeng Sai Srinivas Panapakkam Giridharan Naava Naslavsky Steve Caplan Department of Biochemistry and Molecular Biology and Eppley Cancer CenterUniversity of Nebraska Medical CenterOmahaNE 68198-5870United States

The internalization of essential nutrients,lipids and receptors is a crucial process for all eukaryotic ***,endocytosis is highly conserved across cell types and *** internalized,small cargocontaining vesicles fuse with early endosomes(also known as sorting endosomes),where they undergo segregation to distinct membrane regions and are sorted and transported on through the endocytic *** the mechanisms that regulate this sorting are still poorly understood,some receptors are directed to late endosomes and lysosomes for degradation,whereas other receptors are recycled back to the plasma membrane;either directly or through recycling *** Rab family of small GTP-binding proteins plays crucial roles in regulating these trafficking *** cycle from inactive GDP-bound cytoplasmic proteins to active GTP-bound membraneassociated proteins,as a consequence of the activity of multiple specific GTPase-activating proteins(GAPs) and GTP exchange factors(GEFs).Once bound to GTP,Rabs interact with a multitude of effector proteins that carry out Rab-specific *** studies have shown that some of these effectors are also interaction partners for the C-terminal Eps15 homology(EHD) proteins,which are also intimately involved in endocytic regulation.A particularly interesting example of common Rab-EHD interaction partners is the MICALlike protein,***-L1 and its homolog,MICAL-L2,belong to the larger MICAL family of proteins,and both have been directly implicated in regulating endocytic recycling of cell surface receptors and junctional proteins,as well as controlling cytoskeletal rearrangement and neurite *** this review,we summarize the functional roles of MICAL and Rab proteins,and focus on the significance of their interactions and the implications for endocytic transport.

关键词： Rab MICAL Eps15 homology endosomes Endocytosis Trafficking Cytoskeleton

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