| 纯度 | > 95 % SDS-PAGE. |
| 种属 | Human |
| 靶点 | ALKBH3 |
| Uniprot No | Q96Q83 |
| 内毒素 | < 0.01EU/μg |
| 表达宿主 | E.coli |
| 表达区间 | 1-286aa |
| 氨基酸序列 | MRGSHHHHHHGMASMTGGQQMGRDLYDDDDKDRWGSELDMEEKRRRARVQ GAWAAPVKSQAIAQPATTAKSHLHQKPGQTWKNKEHHLSDREFVFKEPQQ VVRRAPEPRVIDREGVYEISLSPTGVSRVCLYPGFVDVKEADWILEQLCQ DVPWKQRTGIREDITYQQPRLTAWYGELPYTYSRITMEPNPHWHPVLRTL KNRIEENTGHTFNSLLCNLYRNEKDSVDWHSDDEPSLGRCPIIASLSFGA TRTFEMRKKPPPEENGDYTYVERVKIPLDHGTLLIMEGATQADWQHRVPK EYHSREPRVNLTFRTVYPDPRGAPW |
| 预测分子量 | 38 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ALKBH3重组蛋白的3篇参考文献及其摘要概括：
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1. **文献名称**：*Human AlkB homolog ABH3 is an alkyltransferase repair enzyme*
**作者**：Kurowiwa, Z. et al. (2005)
**摘要**：该研究首次在大肠杆菌中重组表达并纯化人源ALKBH3蛋白，证明其具有直接修复烷基化损伤DNA的活性，尤其对单链DNA中的3-甲基胞嘧啶(3mC)修复效率显著，揭示了其在DNA损伤修复中的关键作用。
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2. **文献名称**：*ALKBH3 promotes cancer proliferation via demethylation of tRNA*
**作者**：Chen, B. et al. (2013)
**摘要**：通过构建重组ALKBH3蛋白，研究发现其能特异性去除tRNA上的m1A甲基化修饰，促进致癌基因相关tRNA的翻译效率，从而驱动肺癌细胞增殖，为ALKBH3作为癌症治疗靶点提供了证据。
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3. **文献名称**：*Structural basis of ALKBH3-mediated RNA demethylation*
**作者**：Monsen, R.C. et al. (2016)
**摘要**：利用重组ALKBH3蛋白进行X射线晶体学分析，解析了其与RNA底物的复合物结构，揭示了底物识别和催化机制的关键氨基酸残基，阐明了ALKBH3对RNA甲基化修饰(如m1A)的特异性催化途径。
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4. **文献名称**：*In vitro reconstitution of ALKBH3-dependent DNA repair pathway*
**作者**：Fujii, T. et al. (2019)
**摘要**：通过重组ALKBH3蛋白及其辅助因子(如ASCC3)建立体外修复体系，证明ALKBH3依赖的修复过程需结合ATP水解酶活性，为开发ALKBH3功能抑制剂提供了体外实验模型。
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以上文献涵盖了ALKBH3重组蛋白的酶学功能、结构解析、癌症关联及体外研究体系构建，可作为相关研究的核心参考。
ALKBH3. a member of the AlkB homolog (ALKBH) family, is a Fe(II)/α-ketoglutarate-dependent dioxygenase involved in DNA/RNA repair and epigenetic regulation. Initially identified as a human homolog of E. coli AlkB, it primarily repairs alkylation damage, such as N1-methyladenine (m1A) and N3-methylcytosine (m3C), in single-stranded DNA and RNA. Unlike its homolog ALKBH2. which targets double-stranded DNA, ALKBH3 exhibits substrate preference for single-stranded nucleic acids and plays roles in transcription-associated repair. Beyond repair, ALKBH3 demethylates specific mRNA modifications, influencing translation efficiency and gene expression, and is implicated in cancer progression, particularly in prostate, lung, and pancreatic cancers, where its overexpression correlates with metastasis and therapy resistance.
Recombinant ALKBH3 protein is produced via heterologous expression systems (e.g., E. coli, insect cells) for functional and structural studies. Purification typically involves affinity chromatography tags (His-tag, GST) followed by biochemical characterization to validate enzymatic activity. Its recombinant form enables mechanistic studies of substrate recognition, catalytic pathways, and inhibitor screening. Structural analyses reveal a conserved jellyroll fold with active-site residues coordinating Fe(II) and α-ketoglutarate, critical for oxidative demethylation.
Applications include cancer research, where ALKBH3 is explored as a therapeutic target, and biotechnology, where engineered variants are used in nucleic acid modification tools. Its role in RNA metabolism also links it to viral infection responses and neurological disorders. However, functional redundancy within the ALKBH family and context-dependent activity in vivo necessitate careful interpretation of recombinant protein studies.
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