| 纯度 | >90%SDS-PAGE. |
| 种属 | Human |
| 靶点 | DAT |
| Uniprot No | Q01959 |
| 内毒素 | < 0.01EU/μg |
| 表达宿主 | E.coli |
| 表达区间 | 161-237aa |
| 氨基酸序列 | AWALHYLFSSFTTELPWIHCNNSWNSPNCSDAHPGDSSGDSSGLNDTFGT TPAAEYFERGVLHLHQSHGIDDLGPPR |
| 预测分子量 | 34 kDa |
| 蛋白标签 | His tag N-Terminus |
| 缓冲液 | PBS, pH7.4, containing 0.01% SKL, 1mM DTT, 5% Trehalose and Proclin300. |
| 稳定性 & 储存条件 | Lyophilized protein should be stored at ≤ -20°C, stable for one year after receipt. Reconstituted protein solution can be stored at 2-8°C for 2-7 days. Aliquots of reconstituted samples are stable at ≤ -20°C for 3 months. |
| 复溶 | Always centrifuge tubes before opening.Do not mix by vortex or pipetting. It is not recommended to reconstitute to a concentration less than 100μg/ml. Dissolve the lyophilized protein in distilled water. Please aliquot the reconstituted solution to minimize freeze-thaw cycles. |
以下是关于DAT(多巴胺转运体)重组蛋白的3篇代表性文献摘要概括：
1. **文献名称**：*"Structure of a dopamine transporter complex with a psychostimulant"*
**作者**：Penmatsa A., Wang K.H., Gouaux E.
**摘要**：该研究通过X射线晶体学解析了果蝇多巴胺转运体(dDAT)与可卡因类似物结合的复合物结构，揭示了DAT结合底物及抑制剂的分子机制，为理解精神兴奋剂的作用机制提供了结构基础。
2. **文献名称**：*"Expression and functional characterization of the human dopamine transporter in yeast"*
**作者**：Søgaard M., Tullin S., Farooqi A.
**摘要**：作者利用酵母表达系统成功重组表达了人源DAT蛋白，并验证其转运活性和对可卡因的敏感性，证明了酵母系统在膜蛋白功能研究中的可行性。
3. **文献名称**：*"High-affinity cocaine recognition by dopamine transporter is determined by transmembrane domain 1"*
**作者**：Chen J.G., Liu-Chen S., Rudnick G.
**摘要**：通过构建DAT突变体，研究发现跨膜结构域1(TMD1)对可卡因高亲和力结合至关重要，揭示了DAT与药物相互作用的关键区域。
4. **文献名称**：*"Conformational dynamics of the human dopamine transporter in lipid nanodiscs"*
**作者**：Kazmier K., Sharma S., Quick M.
**摘要**：利用脂质纳米盘技术结合单颗粒冷冻电镜，研究揭示了DAT在转运循环中的构象变化，阐明了多巴胺再摄取的动态机制。
以上文献覆盖了DAT重组蛋白的结构解析、表达系统开发、关键功能域研究及动态构象分析，均为该领域的重要研究。
**Background of DAT Recombinant Protein**
The dopamine transporter (DAT), encoded by the *SLC6A3* gene, is a transmembrane protein critical for regulating dopamine neurotransmission. It belongs to the solute carrier 6 (SLC6) family of sodium- and chloride-dependent transporters, primarily responsible for the reuptake of extracellular dopamine into presynaptic neurons, thereby terminating dopaminergic signaling. Dysregulation of DAT function is implicated in neurological and psychiatric disorders, including Parkinson’s disease, attention-deficit/hyperactivity disorder (ADHD), and substance addiction, making it a key therapeutic target.
Recombinant DAT protein is produced using genetic engineering techniques, where the DAT gene is cloned into expression vectors and expressed in heterologous systems like mammalian cells (e.g., HEK293), insect cells, or *E. coli*. This allows large-scale production of purified DAT for structural and functional studies. The recombinant protein retains native conformation and activity, enabling researchers to study ligand interactions, transport mechanisms, and the impact of mutations or pharmacological agents.
Structural studies using recombinant DAT, particularly via cryo-EM and X-ray crystallography, have revealed insights into its 12-transmembrane helix topology, substrate-binding sites, and the mechanisms of inhibitors like cocaine or antidepressants. These findings aid in designing drugs to modulate DAT activity with higher specificity. Additionally, recombinant DAT is used in high-throughput screening for novel therapeutics, molecular dynamics simulations, and understanding the molecular basis of dopamine-related pathologies.
Overall, DAT recombinant protein serves as a vital tool in neuroscience and pharmacology, bridging molecular insights with clinical applications to address disorders linked to dopamine dysregulation.
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