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Val 70, Phe 72 and the last seven amino acid residues of C-terminal are essential to the function of norepinephrine transporter

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Cell Research 1998年第4期8卷 311-315页

作者： LIU YAN HONG FANG HUANG JIAN FEI JIAN XING ZHAO QUANG BAO GU WOLFGANG SCHWARZ LI HE GUO (Shanghai Institute of Cell Biology, Chinese Academy of Sciences, Shanghai 200031, China)(Max-Plank Institute for Biophysik, Kennedy allee 70,60596 Frankfurt/M G Shanghai Institute of Cell Biology Chinese Academy of Sciences Shanghai China Max-Plank Institute for Biophysik Frankfurt/M Germany

The norepinephrine transporter(NET) is a member of the Na+/Cl- dependent neurotransmitter transporter family and constitutes the target of several clinically important antidepressants. To delineate the critical amino acid residues and the function of C-terminal in regulating transport activity of NET, here we constructed two site mutants (V70F, F72V; V70I, F72V) and one C-terminal truncated mutant (△611-617). The wild type and mutants of NET were expressed in Xenopus oocytes by injection of their cRNA. We found that all of these mutants lost their transport activity. These results indicate that the amino acid residues of V70 and F72 3 and the last seven amino acids of C-terminal are essential to the transport activity of NET.

关键词： Norepinephrine transporter neurotransmitter uptake mutagenesis

Further study on Ca_(2+) -mediated inhibition of human erythrocyte D-glucose transporter

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Chinese Science Bulletin 1997年第10期42卷 871-875页

作者： XIE Wensheng, TU Yaping and YANG FuyuState Key Laboratory of Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, Beijing 100101, China State Key Laboratory of Biomacromolecules Institute of Biophysics Chinese Academy of Sciences Beijing China

THE transport of D-glucose to erythrocytes is catalyzed by an integral membrane protein,i.***.the D-glucose transporter.It specifically carries D-glucose,driven by glucose’s concentrationgradient.Because of its relative abundance and higher transport activity,human erythrocyteglucose transporter has been generally used as a model system to study sugar transport,but

关键词： Ca2+ glucose transporter transporting activity inhibition.

Research of 1,3-Dihydroxyacetone Production by Overexpressing Glycerol transporter and Glycerol Dehydrogenase

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Transactions of Tianjin University 2019年第5期25卷 549-558页

作者： Jinxiu Tan Xiaona Yang Wenyu Lu School of Chemical Engineering and Technology Tianjin UniversityTianjin 300072China Key Laboratory of System Bioengineering (Tianjin University) Ministry of EducationTianjin 300072China Collaborative Innovation Center of Chemical Science and Engineering (Tianjin) SynBio Research PlatformTianjin 300072China

1,3-Dihydroxyacetone (DHA), a natural ketose, is widely used in the chemical, cosmetic, and pharmaceutical industries. The current method for DHA production is Gluconobacter oxydans ( G. oxydans ) fermentation, but the high concentration of glycerol in the fermentation broth inhibits cells growth. To overcome this obstacle, in this study, we overexpressed the glycerol transporter (GlpFp) by the use of promoters P tufB , P gmr , P glp1 , and P glp2 in G. oxydans 621H. The results show that the glycerol tolerances of strains overexpressing G lpF were all much better than that of the control strain. The glycerol dehydrogenase gene (G dh) was overexpressed by the promoters P tufB and P gdh , which increased the DHA titer by 12.7% compared with that of the control group. When G lpF and Gdh genes were co-overexpressed in G. oxydans 621H, the OD600 value of the engineered strains all increased, but the DHA titers decreased in di erent degrees, as compared with strains that overexpressed only G dh . This study provides a reference for future research on DHA production.

关键词： G. oxydans 621H 1,3-dihydroxyacetone Glycerol Glycerol transporter Glycerol dehydrogenase

5-Fluorocytosine–Sugar Conjugates for Glucose transporter-Mediated Tumor Targeting: Synthesis, Cytotoxicity, and Cellular Uptake Mechanism

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Transactions of Tianjin University 2019年第6期25卷 611-617页

作者： Yu Wang Xiaofei Cheng Hongxia Zhao Qingzhi Gao Tianjin Key Laboratory for Modern Drug Delivery and High-Efficiency School of Pharmaceutical Science and Technology Tianjin University

Two novel sugar-conjugated 5-fluorocytosine(5-FC)antineoplastic compounds were designed and synthesized to improve the selective drug uptake by targeting the tumor-specific glucose transporter(GLUT).The antitumor activity of these compounds was evaluated in four different human cancer cell lines:A549(human lung cancer cell line),HT29(human colorectal cancer cell line),H460(human lung cancer cell line),and PC3(human prostate cancer cell line).The sugar conjugates exhibited cytotoxicity similar to or higher than 5-FC and 1-hexylcarbamoyl-5-FC in A549,HT29,H460,and PC3.Furthermore,GLUT-mediated transport of the glycoconjugate was investigated with GLUT inhibitor-mediated cytotoxicity analysis in a GLUT-overexpressing HT29 cell line.The cell-killing potency of 5-FC glycoconjugate was found to depend significantly on the GLUT inhibitor,and the cellular uptake of molecules was regulated by GLUT-mediated transport.All the results demonstrate the potential advantages of glycoconjugation for Warburg effect-targeted drug design.

关键词： Warburg effect Glucose transporter overexpressed 5-Fluorocytosine glycoconjugate Tumor targeting

Human norepinephrine transporter gene

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Chinese Science Bulletin 1995年第17期40卷 1465-1468页

作者：郭礼和朱丽华黄芳 AntnonyC.W.Tam 叶增产费俭张孝勇 DominicMan-KitLam Shanghai Institute of Cell Biology Chinese Academy of Sciences Shanghai 200031 China)

Norepinephrine (NE) is a catecholamine neurotransmitter in postganglionic sympatheticnerves and within the central nervous system. In noradrenergic neurons, synaptictransmission includes three stepst release of NE into the synaptic cleft, interaction with apostsynaptic receptor, and subsequent removal from the cleft into pre-synaptic terminals or glialcells. This reaccumulation process is catalyzed by sodium-dependent NE transporter (NET).

关键词： human norepinephrine transporter gene cloning.

GhKT2: a novel K^+ transporter gene in cotton(Gossypium hirsutum)

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Frontiers of Agricultural Science and Engineering 2018年第2期5卷 226-235页

作者： Yiru WANG Juan XU Mingcai ZHANG Xiaoli TIAN Zhaohu LI Department of Agronomy College of Agronomy and Biotechnology China Agricultural University

Potassium is an essential nutrient for plant growth and productivity of crops. K+transporters are important for K+uptake and transport in plants. However,information on the function of K+transporters and K+channels in cotton is limited. The KT/KUP/HAK protein family is essential for a variety of physiological processes in plants, including nutrient acquisition and regulation of development. This study, identified a K+transporter gene,Gh KT2, expressed in the roots of cotton(Gossypium hirsutum) cv. Liaomian17. The deduced transcript of Gh KT2 is highly homologous to Cluster II of KUP/HAK/KT K+transporters and is predicted to contain 11 transmembrane domains. Gh KT2 has been localized to the plasma membrane, and its transcripts were detected in roots, stems, leaves and shoot apices of cotton seedlings.Consistently, b-glucuronidase(GUS) expression driven by the Gh KT2 promoter could be detected in roots, mesophyll cells, and leaf veins in transgenic Arabidopsis. In addition,the expression of Gh KT2 was induced by low K+stress in cotton roots and p Gh KT2::GUS-transgenic Arabidopsis seedlings. The Gh KT2-overexpression Arabidopsis lines plants were larger and showed greater K+accumulation than the wild type(WT) regardless of K+concentration supplied. The net K+influx rate, measured by the noninvasive micro-test technique, in root meristem zone of Gh KT2-transgenic Arabidopsis lines was significantly greater than that of WT. Taken together, this evidence indicates that Gh KT2 may participate in K+acquisition from low or high external K+, as well as K+transport and distribution in plants.

关键词： cotton GhKT2 potassium transporter uptake

Predicting disruptions to drug pharmacokinetics and the risk of adverse drug reactions in nonalcoholic steatohepatitis patients

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Acta Pharmaceutica Sinica B 2023年第1期13卷 1-28页

作者： Solène Marie Kayla L.Frost Raymond K.Hau Lucy Martinez-Guerrero Jailyn M.Izu Cassandra M.Myers Stephen H.Wright Nathan J.Cherrington College of Pharmacy Department of Pharmacology&ToxicologyUniversity of ArizonaTucsonAZ 85721USA College of Medicine Department of PhysiologyUniversity of ArizonaTucsonAZ 85724USA

The liver plays a central role in the pharmacokinetics of drugs through drug metabolizing enzymes and transporters.Non-alcoholic steatohepatitis(NASH)causes disease-specific alterations to the absorption,distribution,metabolism,and excretion(ADME)processes,including a decrease in protein expression of basolateral uptake transporters,an increase in efflux transporters,and modifications to enzyme activity.This can result in increased drug exposure and adverse drug reactions(ADRs).Our goal was to predict drugs that pose increased risks for ADRs in NASH patients.Bibliographic research identified 71 drugs with reported ADRs in patients with liver disease,mainly non-alcoholic fatty liver disease(NAFLD),54 of which are known substrates of transporters and/or metabolizing enzymes.Since NASH is the progressive form of NAFLD but is most frequently undiagnosed,we identified other drugs at risk based on NASH-specific alterations to ADME processes.Here,we present another list of 71 drugs at risk of pharmacokinetic disruption in NASH,based on their transport and/or metabolism processes.It encompasses drugs from various pharmacological classes for which ADRs may occur when used in NASH patients,especially when eliminated through multiple pathways altered by the disease.Therefore,these results may inform clinicians regarding the selection of drugs for use in NASH patients.

关键词： Non-alcoholic steatohepatitis Hepatic elimination transporter Enzyme Pharmacokinetics Drug-drug interaction Liver Adverse drug reaction

The upregulated intestinal folate transporters direct the uptake of ligand-modified nanoparticles for enhanced oral insulin delivery

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Acta Pharmaceutica Sinica B 2022年第3期12卷 1460-1472页

作者： Jingyi Li Yaqi Zhang Miaorong Yu Aohua Wang Yu Qiu Weiwei Fan Lars Hovgaard Mingshi Yang Yiming Li Rui Wang Xiuying Li Yong Gan School of Pharmacy Shanghai University of Traditional Chinese MedicineShanghai 201203China State Key Laboratory of Drug Research Shanghai Institute of Materia MedicaChinese Academy of SciencesShanghai 201203China University of Chinese Academy of Sciences Beijing 100049China Oral Formulation Development Novo Nordisk A/SMacilov 2760Denmark Department of Pharmacy Faculty of Health and Medical SciencesUniversity of CopenhagenCopenhagen 2100Denmark University of Texas at Dallas RichardsonTX 75080USA NMPA Key Laboratory for Quality Research and Evaluation of Pharmaceutical Excipients National Institutes for Food and Drug ControlBeijing 100050China

transporters are traditionally considered to transport small molecules rather than large-sized nanoparticles due to their small pores.In this study,we demonstrate that the upregulated intestinal transporter(PCFT),which reaches a maximum of 12.3-fold expression in the intestinal epithelial cells of diabetic rats,mediates the uptake of the folic acid-grafted nanoparticles(FNP).Specifically,the upregulated PCFT could exert its function to mediate the endocytosis of FNP and efficiently stimulate the traverse of FNP across enterocytes by the lysosome-evading pathway,Golgi-targeting pathway and basolateral exocytosis,featuring a high oral insulin bioavailability of 14.4%in the diabetic rats.Conversely,in cells with relatively low PCFT expression,the positive surface charge contributes to the cellular uptake of FNP,and FNP are mainly degraded in the lysosomes.Overall,we emphasize that the upregulated intestinal transporters could direct the uptake of ligand-modified nanoparticles by mediating the endocytosis and intracellular trafficking of ligand-modified nanoparticles via the transporter-mediated pathway.This study may also theoretically provide insightful guidelines for the rational design of transporter-targeted nanoparticles to achieve efficient drug delivery in diverse diseases.

关键词： Ligand-modified nanoparticles transporter Proton-coupled folate transporter Expression level Endocytosis Intracellular trafficking Diabetes Oral insulin delivery

Rapid responses:Receptor-like kinases directly regulate the functions of membrane transport proteins in plants

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Journal of Integrative Plant Biology 2022年第7期64卷 1303-1309页

作者： Xiaopeng Li Jingjie Zhang Hongyong Shi Bo Li Jia Li Ministry of Education Key Laboratory of Cell Activities and Stress Adaptations School of Life SciencesLanzhou UniversityLanzhou730000China Guangdong Provincial Key Laboratory of Plant Adaptation and Molecular Design School of Life SciencesGuangzhou UniversityGuangzhou 510006China

Receptor-like kinases(RLKs)are a large group of plant-specific transmembrane proteins mainly acting as receptors or co-receptors of various extracellular signals.They usually turn extracellular signals into intracellular responses via altering gene expression profiles.However,recent studies confirmed that many RLKs can physically interact with diverse membrane-localized transport proteins and regulate their activities for speedy responses in limited tissues or cells.In this minireview,we highlight recent discoveries regarding how RLKs can work with membrane transport proteins collaboratively and thereby trigger cellular responses in a precise and rapid manner.It is anticipated that such regulation broadly presents in plants and more examples will be gradually revealed when in-depth analyses are conducted for the functions of RLKs.

关键词： channel pump receptor-like kinase signal trans-duction transporter