5—羟色胺转运体研究进展

５－羟色胺（５－ＨＴ）是中枢及外周神经系统中一种重要的神经递质,具有多种生物活活性。５－羟色胺转运体（５－ＨＴＴ）可将５－ＨＴ重摄取,降低胞外５－ＨＴ浓度。近来发现５－ＨＴＴ功能异常与某些精神疾病和可卡因成瘾性有关,因此开发对５－ＨＴＴ具有高亲和力与高选择性的药物在临床上很有意义。目前５－ＨＴＴ已在大鼠和人中克隆,这不仅有助于阐明配体与５－ＨＴＴ作用的分子机制,同时也为药物设计提供了方向。     

高亲和力谷氨酸转运体

高亲和力谷氨酸转运体主要位于神经元和胶质细胞的细胞膜上,能逆浓度梯度从胞外向胞内摄取谷氨酸,中止谷氨酸能传递,使胞外谷氨酸浓度保持在较低水平,以保护神经元不受谷氨酸的毒性影响。近年来,随着高亲和力谷氨酸转运体的克隆,有关研究迅速发展。本文从高亲和力谷氨酸转运体的克隆、分子结构特征、表达分布、生理功能、结构－功能关系等方面对近年的进展加以综述。     

甘氨酸神经递质研究进展

甘氨酸是化学结构最简单的氨基酸,但具有复杂的功能。甘氨酸在中枢神经系统中是介导快速抑制性神经传递的一种重要的神经递质,在控制神经元兴奋性方面发挥重要作用。就其神经递质功能对甘氨酸的生物合成、释放与调控以及作用模式等方面的近年研究进展做一综述,对甘氨酸神经递质的全面认识将有益于炎性痛、痉挛状态以及癫痫等中枢神经系统疾病的诊断、预防及治疗。     

肾脏尿酸转运体的研究进展

尿酸是人体内嘌呤代谢的终产物。在肝脏合成后,正常情况下约70%的尿酸在肾脏分泌,而一小部分则被分泌到肠内。肾脏是血尿酸稳态调节的主要器官,其调控依赖于尿酸转运体的转运。尿酸代谢紊乱或转运异常将导致高尿酸血症、痛风、痛风性肾结石等疾病。本文对肾脏尿酸转运体的研究进展进行综述。     

去甲肾上腺素转运体的研究进展

随着分子克隆技术的发展和应用,已经可以克隆出人的去甲肾上腺素转运体(Norepinephrine transporter,NET)基因转染进哺乳动物细胞内进行体外研究。去甲肾上腺素转运体在神经传递中有着非常重要的作用,许多神经以及精神系统方面的疾病,心血管疾病等都与去甲肾上腺素转运体的功能缺失或紊乱有关。本文主要介绍了去甲肾上腺素转运体的基本概念和近年来国外对去甲肾上腺素转运体的研究概况,综述了与该转运体相关的药物的研究进展以及由于去甲肾上腺素转运体的功能紊乱或丧失而导致的疾病的临床研究。最近几年对于NET基因表达调控以及各种临床疾病的研究对这些疾病的治疗方法的探索有着非常重要的意义。     

神经递质转运体

神经递质转运体(neurotransmitter transporters)或称神经转运体(neurotransporters)存在于神经末梢的细胞膜上,其作用是通过重摄取已释放的递质使其作用终止。这些转运体是突触的关键成份之一,并是许多药物(如抗抑郁药物和抗精神病药物)的作用位点。转运体     

植物质膜钾离子转运体研究进展

近年,随着分子生物学技术的不断发展和广泛应用,有关植物质膜钾离子转运体的研究取得重要进展。目前已经克隆到多种质膜钾离子转运体基因并对钾离子转运体生化特性以及结构功能进行广泛研究。研究认为,质膜钾离子转运体可分为钾离子载体和钾离子通道。钾离子通道又可分为内向性Ｋ＾＋通道α亚基、Ｋ＾＋通道β亚基及外生Ｋ＾＋通道等三类。本文对上述质膜钾离子转运体的生化特性以及结构功能研究的进展进行了综述。     

五羟色胺转运体的研究进展

五羟色胺转运体是一种对五羟色胺（5-HT,serotonin）有高度亲和力的跨膜转运蛋白,能够重新摄取细胞间隙内的5-HT,从而调节神经信号的转导。该文简述了五羟色胺转运体的生物学特性、分布以及与人类疾病的关系,通过分析比较发现,五羟色胺转运体的多态性与肠易激综合征、抑郁症、强迫症都有着密切的关系。     

植物质膜钾离子转运体研究进展

近年,随着分子生物学技术的不断发展和广泛应用,有关植物质膜钾离子转运体的研究取得重要进展。目前已经克隆到多种质膜钾离子转运体基因并对钾离子转运体生化特性以及结构功能进行了广泛研究。研究认为,质膜钾离子转运体可分为钾离子载体和钾离子通道。钾离子通道又可分为内向性K+通道α亚基、K+通道β亚基及外向性K+通道等三类。本文对上述质膜钾离子转运体的生化特性以及结构功能研究的进展进行了综述。     

帕金森病基底神经节神经递质失衡的研究进展

帕金森病(Parkinson's disease,PD)是一种老年神经系统退变性疾病,主要病理改变是中脑黑质致密部多巴胺能神经元渐进性变性死亡,从而引起基底神经节的功能失调。近年的研究显示,多巴胺能神经元的丢失并不是帕金森病发病的唯一因素,基底神经节中其它神经递质,包括谷氨酸、γ-氨基丁酸、乙酰胆碱等,在帕金森病的发病中也有重要的作用。在疾病发生发展过程中,神经递质的合成、分泌发生紊乱,基底神经节网络调控功能失调,导致了以运动系统症状为主的临床表现。本文就帕金森病状态下基底神经节中主要神经递质失衡的研究进展作一综述。     

A comparative computational analysis of protein sequences and literature mining classify 'orphan' neurotransmitter transporters

Various sources of protein data, such as knowledgebases and scientific literature, are currently available, as are numerous tools for their analysis. The matter becomes one of choosing the tools that are most appropriate for the specific task and for the specific proteins. A combination of standard and alternative tools may lead to biologically significant results.Here, a computational classification of proteins is made using standard multiple sequence alignment in combination with an alternative method for analysis of hydropathy distribution in proteins. Both of these methods are applied to the Na⁺/Cl⁻-dependent neurotransmitter symporters (NSSs), resulting in two alternative classifications. The classifications are validated and interpreted biologically by literature and knowledgebase annotation mining, producing a consensus classification. The classification leads to the identification and functional characterization of three families of largely structurally and functionally uncharacterized orphan NSSs. The literature and knowledgebase annotations are mined to functionally characterize the NSSs in these families. The presented work also demonstrates that, in specific cases, the analysis of the hydropathy distribution in proteins is capable of revealing functional properties of proteins.     

Vesicular neurotransmitter transporters

Neurotransmission depends on the regulated release of chemical transmitter molecules. This requires the packaging of these substances into the specialized secretory vesicles of neurons and neuroendocrine cells, a process mediated by specific vesicular transporters. The family of genes encoding the vesicular transporters for biogenic amines and acetylcholine have recently been cloned. Direct comparison of their transport characteristics and pharmacology provides information about vesicular transport bioenergetics, substrate feature recognition by each transporter, and the role of vesicular amine storage in the mechanism of action of psychopharmacologic and neurotoxic agents. Regulation of vesicular transport activity may affect levels of neurotransmitter available for neurosecretion and be an important site for the regulation of synaptic function. Gene knockout studies have determined vesicular transport function is critical for survival and have enabled further evaluation of the role of vesicular neurotransmitter transporters in behavior and neurotoxicity. Molecular analysis is beginning to reveal the sites involved in vesicular transporter function and the sites that determine substrate specificity. In addition, the molecular basis for the selective targeting of these transporters to specific vesicle populations and the biogenesis of monoaminergic and cholinergic synaptic vesicles are areas of research that are currently being explored. This information provides new insights into the pharmacology and physiology of biogenic amine and acetylcholine vesicular storage in cardiovascular, endocrine, and central nervous system function and has important implications for neurodegenerative disease.     

Structure and localisation of drug binding sites on neurotransmitter transporters

The dopamine (DAT), serotontin (SERT) and noradrenalin (NET) transporters are molecular targets for different classes of psychotropic drugs. The crystal structure of Aquifex aeolicus LeuT_Aa was used as a template for molecular modeling of DAT, SERT and NET, and two putative drug binding sites (pocket 1 and 2) in each transporter were identified. Cocaine was docked into binding pocket 1 of DAT, corresponding to the leucine binding site in LeuT_Aa, which involved transmembrane helices (TMHs) 1, 3, 6 and 8. Clomipramine was docked into binding pocket 2 of DAT, involving TMHs 1, 3, 6, 10 and 11, and extracellular loops 4 and 6, corresponding to the clomipramine binding site in a crystal structure of a LeuT_Aa–clomipramine complex. The structures of the proposed cocaine- and tricyclic antidepressant-binding sites may be of particular interest for the design of novel DAT interacting ligands.     

Physiological characteristics of serotonin transporters on rat platelets

Physiological characteristics of serotonin (5-hydroxytryptamine, 5HT) transport through the platelet membrane was investigated in Wistar rats with our recently developed method permitting repetitive measurements of transporter kinetics in individual animals. Full kinetic analysis in the population of 91 animals revealed Michaelis constant (K_m) of 0.158±0.025 μM and maximal velocity (V_max) of 5HT uptake of 225±32 pmol per 10⁸platelets min⁻¹ (mean±S.D.). Both kinetic parameters demonstrated normal distribution curves, which for V_max were slightly skewed toward higher than average values. No gender effect was shown in frequency distributions, mean values and variability of kinetic parameters. A significant intraindividual correlation between kinetic parameters was found suggesting compensation at the level of the plasma membrane. Kinetic parameters were not influenced by age (until the middle age) or annual cycle (under laboratory conditions) and were shown to be fairly stable in time, supporting the view that platelet 5HT transport kinetics could be a useful biological trait marker.     

Using a collection of MUPP1 domains to investigate the similarities of neurotransmitter transporters C-terminal PDZ motifs

A ubiquitous feature of neurotransmitter transporters is the presence of short C-terminal PDZ binding motifs acting as important trafficking elements. Depending on their very C-terminal sequences, PDZ binding motifs are usually divided into at least three groups; however this classification has recently been questioned. To introduce a 3D aspect into transporter’s PDZ motif similarities, we compared their interactions with the natural collection of all 13 PDZ domains of the largest PDZ binding protein MUPP1. The GABA, glycine and serotonin transporters showed unique binding preferences scattered over one or several MUPP1 domains. On the contrary, the dopamine and norepinephrine transporter PDZ motifs did not show any significant affinity to MUPP1 domains. Interestingly, despite their terminal sequence diversity all three GABA transporter PDZ motifs interacted with MUPP1 domain 7. These results indicate that similarities in binding schemes of individual transporter groups might exist. Results also suggest the existence of variable PDZ binding modes, allowing several transporters to interact with identical PDZ domains and potentially share interaction partners in vivo.     

Nitrate transporters and peptide transporters

In higher plants, two types of nitrate transporters, NRT1 and NRT2, have been identified. In Arabidopsis, there are 53 NRT1 genes and 7 NRT2 genes. NRT2 are high-affinity nitrate transporters, while most members of the NRT1 family are low-affinity nitrate transporters. The exception is CHL1 (AtNRT1.1), which is a dual-affinity nitrate transporter, its mode of action being switched by phosphorylation and dephosphorylation of threonine 101. Two of the NRT1 genes, CHL1 and AtNRT1.2, and two of the NRT2 genes, AtNRT2.1 and AtNRT2.2, are known to be involved in nitrate uptake. In addition, AtNRT1.4 is required for petiole nitrate storage. On the other hand, some members of the NRT1 family are dipeptide transporters, called PTRs, which transport a broad spectrum of di/tripeptides. In barley, HvPTR1, expressed in the plasma membrane of scutellar epithelial cells, is involved in mobilizing peptides, produced by hydrolysis of endosperm storage protein, to the developing embryo. In higher plants, there is another family of peptide transporters, called oligopeptide transporters (OPTs), which transport tetra/pentapeptides. In addition, some OPTs transport GSH, GSSH, GSH conjugates, phytochelatins, and metals.     

The evolution fo rhodopsins and neurotransmitter receptors

Summary Rhodopsins share a limited number of amino acid identities with a variety of other integral membrane proteins. Most of these proteins have seven putative transmembrane segments and are likely to play a role in transmembrane signaling. We have undertaken a systematic series of comparisons of primary and secondary structure in order to clarify the functional and evolutionary significance of these sequence similarities. On the basis of consistently high similarity scores, we find that the most internally consistent definition of the rhodopsis gene family would ionclude vertebrate rhodospins, - and -adrenergic receptors, M1 and M2 muscarinic acetylcholine receptors, substance K receptors and insect rhodopsins, while excluding bacterirhodopsin, themas human oncogene, vertebrate and insect nicotinic acetylcholine receptors, and the yeast STE2 and STE3 peptide receptors. The rhodopsin gene family is highly diverged at the primary sequence level but has maintained a conserved secondary structure, including a previosuly unidentified hierarchy of transmembrane segment hydrophobicity. We have deevelope new computer alogithms for progressive multiple sequence alignment and the analysis of local conservation of protein domains, and we have used these algorithms to examined the phylogeny of the rhodopsin gene family and the changing domains of sequence conservation. The results show striking diffiierences and similarities in the conserved domains in each of the three main branches of the rhodopsin gene family, and indicte that color vision arose independently in the lines of descent leading to modern humans and fruit flies.     

Glutathione transporters

Background

Glutathione (GSH) is synthesized in the cytoplasm but there is a requirement for glutathione not only in the cytoplasm, but in the other organelles and the extracellular milieu. GSH is also imported into the cytoplasm. The transports of glutathione across these different membranes in different systems have been biochemically demonstrated. However the molecular identity of the transporters has been established only in a few cases.

Scope of review

An attempt has been made to present the current state of knowledge of glutathione transporters from different organisms as well as different organelles. These include the most well characterized transporters, the yeast high-affinity, high-specificity glutathione transporters involved in import into the cytoplasm, and the mammalian MRP proteins involved in low affinity glutathione efflux from the cytoplasm. Other glutathione transporters that have been described either with direct or indirect evidences are also discussed.

Major conclusions

The molecular identity of a few glutathione transporters has been unambiguously established but there is a need to identify the transporters of other systems and organelles. There is a lack of direct evidence establishing transport by suggested transporters in many cases. Studies with the high affinity transporters have led to important structure-function insights.

General significance

An understanding of glutathione transporters is critical to our understanding of redox homeostasis in living cells. By presenting our current state of understanding and the gaps in our knowledge the review hopes to stimulate research in these fields. This article is part of a Special Issue entitled Cellular functions of glutathione.     

The evolutionary divergence of neurotransmitter receptors and second-messenger pathways

Members of the superfamily of G-protein-coupled neurotransmitter receptors have a conserved secondary structure, a moderate and reasonably steady rate of sequence change, and usually lack introns within the coding sequence. These properties are advantageous for evolutionary studies. The duplication and divergence of the genes in this gene family led to the formation of distinct neurotransmitter pathways and may have facilitated the evolution of complex nervous systems. I have analyzed this evolutionary divergence by quantitative multiple sequence alignment, bootstrap resampling, and statistical analysis of 49 adrenergic, muscarinic cholinergic, dopamine, and octopamine receptor sequences from 12 animal species. The results indicate that the first event to occur within this gene family was the divergence of the catecholamine receptors from the muscarinic acetylcholine receptors, which occurred prior to the divergence of the arthropod and vertebrate lineages. Subsequently, the ability to activate specific second-messenger pathways diverged independently in both the muscarinic and the catecholamine receptors. This appears to have occurred after the divergence of the arthropod and vertebrate lineages but before the divergence of the avian and mammalian lineages. However, the second-messenger pathways activated by adrenergic and dopamine receptors did not diverge independently. Rather, the ability of the catecholamine receptors to bind to specific ligands, such as epinephrine, norepinephrine, dopamine, or octopamine, was repeatedly modified in evolutionary history, and in some cases was modified after the divergence of the second-messenger pathways.     

Analysis of classical neurotransmitter markers in tapeworms: Evidence for extensive loss of neurotransmitter pathways

Parasitic flatworms have complex neuromuscular systems that serve important functions in their life cycles. However, our understanding of neurotransmission in parasitic flatworms is limited. Pioneering studies have suggested the presence of several classical neurotransmitter systems, but their molecular components have not been characterized in most cases. Because these components are conserved in bilaterian animals, we searched the genomes of parasitic flatworms for orthologs of genes required for neurotransmitter synthesis, vesicular transport, reuptake, and reception. Our results indicate that tapeworms have lost the genes that are specifically required in other animals for synaptic signaling using the classical neurotransmitters dopamine, tyramine, octopamine, histamine and gamma-aminobutyric acid (GABA). These results imply that these signaling pathways are either absent in these parasites, or that they require completely different molecular components in comparison with other animals. The orthologs of genes related to histaminergic and GABA signaling are also missing in trematodes (although Schistosoma-specific histaminergic receptors have been previously described). In contrast, conserved genes required for glutamatergic, serotonergic and cholinergic signaling could be found in all analyzed flatworms. We analyzed the expression of selected markers of each pathway in the tapeworm Hymenolepis microstoma by whole-mount in situ hybridization. Each marker was specifically expressed in the nervous system, although with different patterns. In addition, we analyzed the expression of proprotein convertase 2 as a marker of peptidergic cells. This gene showed the widest expression in the nervous system, but was also expressed in other tissues, suggesting additional roles of peptidergic signaling in tapeworm development and reproduction.