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1.
烟碱型乙酰胆碱受体参与乙酰胆碱调控的气孔运动 总被引:2,自引:0,他引:2
动物细胞中 ,乙酰胆碱功能的发挥要求乙酰胆碱受体的参与 ,烟碱型受体的激活剂可以直接影响膜对离子的通透性 .在乙酰胆碱诱导的气孔开放过程中 ,可能同样涉及到烟碱型受体的作用 ,药理学的证据表明烟碱型乙酰胆碱受体参与乙酰胆碱调控的气孔运动 ,而且烟碱型乙酰胆碱受体介导的气孔开放与介质中的离子组成密切相关 ,只有在含K+的介质中烟碱才可以诱导气孔开放而在含Ca2 +的介质中没有作用 ;同样 ,烟碱型乙酰胆碱受体的抑制剂只有在含K+的介质中才能抑制乙酰胆碱诱导的气孔开放 .进一步利用荧光定位技术证明烟碱型受体存在于蚕豆气孔保卫细胞中 ,而且主要分布在保卫细胞原生质体的表面 .免疫印迹实验初步证明在保卫细胞原生质体的微粒体中存在着能与动物烟碱型乙酰胆碱受体的α和β亚基发生免疫交叉反应的蛋白条带 .以上结果表明烟碱型乙酰胆碱受体存在于保卫细胞中 ,而且介导了乙酰胆碱诱导的气孔在含K+介质中的开放 . 相似文献
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肺癌作为当今世界最为致命的疾病之一,严重地威胁着人类的生命健康。神经递质乙酰胆碱(acetylcholine,ACh)作为肺癌的自分泌和旁分泌生长因子,可与肺癌细胞上的烟碱型乙酰胆碱受体(nicotinic acetylcholine receptor,nAChRs)和毒蕈碱型乙酰胆碱受体(muscarinic acetylcholine receptors,mAChRs)结合,促进肺癌细胞的增殖、迁移和侵袭。近年来,全基因组数据研究表明,nAChRs与吸烟相关肺癌风险密切相关,因此其在肺癌发生发展中的作用机制的研究对肺癌药物的研发具有重要意义。本文对乙酰胆碱信号通路在肺癌进展中的作用机制进行综述,为其成为肺癌治疗新分子靶点和药物的开发提供依据。 相似文献
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哺乳动物脑烟碱型乙酰胆碱受体的研究进展(综述) 总被引:2,自引:0,他引:2
汪海 《中国应用生理学杂志》1992,8(2):189-192
哺乳动物烟碱乙酰胆碱型受体(nAChR)丰要分布于神经肌肉接点、自主神经节及脑组织。神经肌肉接点突触后膜nAChR的功能为十烃季胺(C_(10))所竞争性拮抗,称C_(10)型nAChR。自主神经节突触后膜nAChR的功能为六烃季铵(C_6)所竞争性拮抗,称C_6型nAChR。脑nAChR对激动剂烟碱、对拮抗剂C_6、C_(10)的敏感性等药理学性质不同于C_(10)nAChR,也不完全与C_6nAchR相似,是nAChR的一种新亚型。与CmnAChR的研究相比,脑nAChR的研究明 相似文献
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烟碱样乙酰胆碱受体(AChR)是一种配基门控性离子通道,由5个亚单位(α_2βγα)构成。利用非洲蟾蜍卵母细胞的表达系统可以研究AChR的通道特性和各亚单位所起的作用。电鳗电器官AChR和小牛肌AChR之间门控特性的差别,主要是由δ亚单位决定的;而小牛肌成年型AChR与胚胎型AChR之间的差别,则由ε亚单位决定。 相似文献
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烟碱型乙酰胆碱受体(nicotinic acetylcholine receptors, nAChRs)是由5个亚基组合形成的配体门控离子通道。根据亚基组成和排列的不同, nAChRs存在多种亚型。由于结构与乙酰胆碱相似,烟碱也是nAChRs的激动剂。大量研究表明,烟碱能影响nAChRs的表达和活性。同时, nAChRs又介导了烟碱成瘾的机制。文中基于最新的研究成果,简要综述了nAChRs的类型、分布和测定方法,并重点阐述了烟碱调控nAChRs表达的机制以及nAChRs在烟碱成瘾过程中的介导作用。 相似文献
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烟碱型乙酰胆碱受体及其亚单位的结构功能 总被引:5,自引:0,他引:5
烟碱型乙酰胆碱受体是配体门控的离子通道蛋白,每个受体由5个亚单位组成,每个亚单位具有4个跨膜α螺旋结构域。通道孔由5个亚单位的M2结构域(α螺旋)环绕围成,它们放射状地排列,使通道孔在膜中部逐渐变细。另外15个α螺旋相互盘绕,构成通道的外环。至今已确认了17种亚单位,它们由一个共同起源的基因家族编码,在中枢神经系统、周围神经系统和肌肉组织广泛表达。众多的亚单位,再加上组织表达的特异性导致受体类型的多样性,结果使该受体具有广泛的生理、药理特性和功能。进年来,基因敲除等现代遗传操作技术提供了在体内研究各类型受体的有效手段,使人类对该受体及亚单位的结构和功能有了长足的认识。 相似文献
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新近研究证实,神经元烟碱型乙酰胆碱受体(nAChR)激动后可起到一定的神经保护作用.目前,一些作用于烟碱受体的激动剂已被作为治疗神经退行性疾病如阿尔茨海默病(AD)和帕金森病(PD)的候选药物,但是关于烟碱受体激动后如何发挥神经保护作用及其潜在的分子机制还不清楚,其中有与Ca2+相关的信号转导假说以及神经营养因子等假说.本文就烟碱型乙酰胆碱受体及其神经保护作用的研究进展予以综述. 相似文献
8.
纯化眼镜蛇毒蛋白对乙酰胆碱受体通道的阻断作用 总被引:4,自引:0,他引:4
在培养的抓蟾胚胎神经和肌肉细胞上用膜片箝技术研究了从中国华南产眼镜蛇毒分离出的神经毒组分对乙酰胆碱受体(ACh-R)通道的影响,发现它在微终板电位和单通道水平上都有明显的阻断作用。完全阻断神经肌肉接点传递的剂量约为2.0μg/ml,对乙酰胆碱受体通道的半阻断量约为0.23μg/ml。这表明它与α-银环蛇毒有相似功效。 相似文献
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alpha-Galactoside Binding Proteins from Plant Membranes: Isolation, Characterization, and Relation to Helminthosporoside Binding Proteins of Sugarcane
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α-Galactoside binding proteins were isolated from cellular membranes of mint and tobacco as well as two clones of sugarcane which differ in their sensitivity to helminthosporoside, a toxic galactoside. Sodium trichloroacetate was used to disrupt membranes after which the proteins were purified using a melibiose-Sepharose-6B affinity column. Proteins from mint, tobacco, and susceptible sugarcane had equal electrophoretic mobilities, whereas resistant sugarcane protein migrated more slowly. Pretreatment of the proteins with fluorescamine caused them to migrate with the tracking dye. Each of the proteins had molecular weights of about 100,000 and each was shown to be oligomeric. Gel filtration revealed that aqueous solutions of these membrane proteins contained a mixture of size species which included a high molecular weight multimer and lower molecular weight oligomers. The relative abundance of the oligomers was dependent upon protein concentration: the lower concentrations yielded higher relative amounts of oligomers (Kenfield and Strobel 1980 Biochim Biophys Acta 600: 705-712). Also, the binding activity of these receptors was inversely proportional to protein concentration. At low protein concentration (4 micrograms per milliliter), the Kd's of each of the proteins for galactinol, raffinose, and helminthosporoside was about 10 micromolar. At high protein concentrations (100 micrograms per milliliter), mint and resistant sugarcane proteins failed to bind α-galactosyl ligands, whereas proteins from tobacco and susceptible sugarcane exhibited a markedly decreased binding activity compared to that at lower protein concentrations. Binding proteins from susceptible sugarcane were mixed with receptors from either resistant sugarcane or mint at low protein concentrations, then assayed for binding activity. Such mixtures showed a concentration-dependent decrease in binding activity analogous to the activity of homogeneous protein solutions. Bovine serum albumin, a nonsubunit protein, had no effect on the binding activity of the protein from susceptible sugarcane. Thus, receptors from diverse plants can associate in vitro and form functional oligomers. The amino acid composition of each of the binding proteins was similar but not identical. The significance of these results is discussed in regard to regulation of carbohydrate transport and sensitivity to phytotoxins. 相似文献
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氧化固醇结合蛋白结构、功能与应用 总被引:1,自引:0,他引:1
氧化固醇结合蛋白(oxysterol binding protein,OSBP)是存在于真核细胞内的一类参与脂质代谢的非囊泡运输蛋白质,在哺乳动物中被称为氧化固醇结合蛋白相关蛋白质(oxysterol binding protein-related proteins,ORPs),而在酵母中被称为氧化固醇结合蛋白同源物质(oxysterol-binding protein homologues,OSH)。近年来人们对氧化固醇结合蛋白的研究不断深入,特别是对其同源蛋白质(例如,ORP5/8、Osh3/4、ORP4L等)的结构功能差异和其在信号转导中的作用的相关研究,以及在生物医药方面的应用更成为了本领域的热点。本文综述了关于OSBP及其同源蛋白质结构和功能的相关研究,指出了该领域存在的一些关键问题。与此同时,对OSH和ORPs在细胞内的膜接触位点(membrane contact sites,MCS)进行对比,以及对今后OSBP的研究方向做了展望。 相似文献
14.
Riaz Mahmood Bidyottam Mittra Jane C. Hines Dan S. Ray 《Molecular and cellular biology》2001,21(14):4453-4459
The Crithidia fasciculata cycling sequence binding protein (CSBP) binds with high specificity to sequence elements in several mRNAs that accumulate periodically during the cell cycle. Mutations in these sequence elements abolish both cycling of the mRNA and binding of CSBP. Two genes, CSBPA and CSBPB, encoding putative subunits of CSBP have been cloned and were found to be present in tandem on the same DNA molecule and to be closely related. CSBPA and CSBPB are predicted to encode proteins with sizes of 35.6 and 42.0 kDa, respectively. Both CSBPA and CSBPB proteins have a predicted coiled-coil domain near the N terminus and a novel histidine and cysteine motif near the C terminus. The latter motif is conserved in other trypanosomatid species. Gel sieving chromatography and glycerol gradient sedimentation results indicate that CSBP has a molecular mass in excess of 200 kDa and an extended structure. Recombinant CSBPA and CSBPB also bind specifically to the cycling sequence and together can be reconstituted to give an RNA gel shift similar to that of purified CSBP. Proteins in cell extracts bind to an RNA probe containing six copies of the cycling sequence. The RNA-protein complexes contain both CSBPA and CSBPB, and the binding activity cycles in near synchrony with target mRNA levels. CSBPA and CSBPB mRNA and protein levels show little variation throughout the cell cycle, suggesting that additional factors are involved in the cyclic binding to the cycling sequence elements. 相似文献
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By means of differential centrifugation, cytosol fraction and microsome were prepared from maize roots which have been grown in dark for 4 d. Highly purified plasma membranes were isolated from the microsome in two-phase aqueous system which is composed of 6.9 % (W/W) Dextran T500 and PEG 3350. The tonoplast was collected from the interface between 1% and 8% (W/W) Dextran T70 after gradient centrifugation. Electron microscopic observation and marker enzyme activities analysis proved that these fractions contained very few other membranes. Microvolume radioactivity ligand binding assay indicated that the specific binding sites of ABA in maize root microsome were mainly distributed on tonoplast and plasma membrane fractions. Their specific binding activity was 2485.4 and 1257.3 fmol/mg protein, respectively, the specific binding activity of cytosol fraction being the lowest (one order of magnitude lower). The dissociation constant (KD) of ABA-BP in plasma membrane was 1.57 nmol/L. 相似文献
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Sean C. Myers-Payne Timothy Hubbell Lixia Pu Frank Schnütgen Torsten Börchers †W. Gibson Wood Friedrich Spener Friedhelm Schroeder 《Journal of neurochemistry》1996,66(4):1648-1656
Abstract: Two fatty acid binding proteins (FABPs) were isolated from Swiss Webster mouse brains. Neither protein cross-reacted with antisera to recombinant liver L-FABP. One protein, designated brain H-FABP, migrated on tricine sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) as a single band at 14.5 kDa with pl 4.9. Brain H-FABP bound NBD-stearic acid and cis -parinaric acid with K D values near 0.02 and 0.5 µ M , respectively. Brain H-FABP cross-reacted with affinity-purified antisera to recombinant heart H-FABP. The second protein, mouse brain B-FABP, migrated on tricine SDS-PAGE gels as a doublet at 16.0 and 15.5 kDa with pl values of 4.5 and 4.7, respectively. Brain B-FABP bound NBD-stearic acid and cis -parinaric acid with K D values near 0.01 and 0.7 µ M , respectively. The brain B-FABP doublet was immunoreactive with affinity-purified antibodies against recombinant mouse brain B-FABP, but not with affinity-purified antibodies against heart H-FABP. [3 H]Oleate competition binding indicated that the two brain FABPs had distinct ligand binding specificities. Both bound fatty acids, fatty acyl CoA, and lysophosphatidic acid. Although both preferentially bound unsaturated fatty acids, twofold differences in specific saturated fatty acid binding were observed. Brain B-FABP and brain H-FABP represented 0.1 and 0.01% of brain total cytosolic protein, respectively. In summary, mouse brain contains two native fatty acid binding proteins, brain H-FABP and brain B-FABP. 相似文献
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PC12 pheochromocytoma cells take up 3,4-dihydroxyphenylethylamine (dopamine) and norepinephrine by a Na+-dependent, cocaine-sensitive system. The kinetics suggest that the same transporter functions for both substrates. Xylamine, a nitrogen mustard that blocks catecholamine uptake into neurons, irreversibly inhibited norepinephrine uptake into PC12 (IC50 = 15 microM). Pretreatment with 10 microM xylamine did not inhibit norepinephrine transport if 10 microM cocaine or 100 microM norepinephrine was also present during the pretreatment period or if Na+ was absent. These results indicate that xylamine must interact with the norepinephrine transporter to inhibit norepinephrine uptake. PC12 accumulated [3H]xylamine; this uptake had Na+-dependent and Na+-independent components. The Na+-dependent uptake was saturable (Km = 13 microM), and it was inhibited by cocaine (IC50 = 0.6 microM), desipramine (IC50 less than 1 nM), and norepinephrine (IC50 = 1 microM). Several proteins became prominently labeled when intact PC12 cells were incubated with [3H]xylamine; these proteins were enriched in a plasma membrane fraction and have molecular weights of 17,000, 24,000, 31,000, 33,000, 41,000, 42,000, 52,000, and 80,000. Other proteins were labeled less prominently. The labeling of all proteins was markedly decreased when the incubation with [3H]xylamine occurred in the presence of cocaine, desipramine, gramicidin D, or in a Na+-free buffer. These results indicate that xylamine must be transported into the cells for covalent binding to proteins to occur. [3H]Xylamine labeled essentially the same proteins when incubated with cell homogenates, but competition experiments with bretylium, desipramine, and cocaine failed to reveal which of the [3H]xylamine-labeled proteins is associated with the norepinephrine transporter. 相似文献
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Karolina Michalska Changsoo Chang Jamey C. Mack Sarah ZerbsAndrzej Joachimiak Frank R. Collart 《Journal of molecular biology》2012,423(4):555-575
In vitro growth experiments have demonstrated that aromatic compounds derived from lignin can be metabolized and represent a major carbon resource for many soil bacteria. However, the proteins that mediate the movement of these metabolites across the cell membrane have not been thoroughly characterized. To address this deficiency, we used a library representative of lignin degradation products and a thermal stability screen to determine ligand specificity for a set of solute-binding proteins (SBPs) from ATP-binding cassette (ABC) transporters. The ligand mapping process identified a set of proteins from Alphaproteobacteria that recognize various benzoate derivatives. Seven high-resolution crystal structures of these proteins in complex with four different aromatic compounds were obtained. The protein-ligand complexes provide details of molecular recognition that can be used to infer binding specificity. This structure-function characterization provides new insight for the biological roles of these ABC transporters and their SBPs, which had been previously annotated as branched-chain amino‐acid-binding proteins. The knowledge derived from the crystal structures provides a foundation for development of sequence-based methods to predict the ligand specificity of other uncharacterized transporters. These results also demonstrate that Alphaproteobacteria possess a diverse set of transport capabilities for lignin-derived compounds. Characterization of this new class of transporters improves genomic annotation projects and provides insight into the metabolic potential of soil bacteria. 相似文献
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Gibberellin-binding proteins were found on the membrane of young rice shoot. The dissociation constant (Kd) for GAs was approximately 6.5 × 10-8 mol/L, and the total concentration of the sites was 0. 3 pmol ·mg-1 protein. The binding activity of gibberellin-binding proteins was significantly affected by temperature and phi which was 140% higher at 0 ℃ than that at 25 ℃, and the optimal pH value was 5. Gibberellin-binding activity increased with the incubation time, reaching the maximum at 1 h. and then decreased gradually. Both IAA and ABA were able to compete with GA3 for gibberellin-binding proteins. 相似文献