纯度 | >90%SDS-PAGE. |
种属 | Human |
靶点 | TK2 |
Uniprot No | O00142 |
内毒素 | < 0.01EU/μg |
表达宿主 | E.coli |
表达区间 | 34-265aa |
氨基酸序列 | VQRRAWPPDKEQEKEKKSVICVEGNIASGKTTCLEFFSNATDVEVLTEPVSKWRNVRGHNPLGLMYHDASRWGLTLQTYVQLTMLDRHTRPQVSSVRLMERSIHSARYIFVENLYRSGKMPEVDYVVLSEWFDWILRNMDVSVDLIVYLRTNPETCYQRLKKRCREEEKVIPLEYLEAIHHLHEEWLIKGSLFPMAAPVLVIEADHHMERMLELFEQNRDRILTPENRKHCP |
预测分子量 | 32.5 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TK2(胸苷激酶2)重组蛋白的虚构参考文献示例(实际文献需通过学术数据库检索):
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1. **文献名称**: *Recombinant Human TK2: Expression, Purification, and Enzymatic Characterization*
**作者**: Wang, X. et al.
**摘要**: 本研究报道了在大肠杆菌中高效表达和纯化重组人源TK2蛋白的方法,并验证其磷酸化胸苷和脱氧胞苷的酶活性,为线粒体疾病研究提供工具。
2. **文献名称**: *AAV-mediated Delivery of Recombinant TK2 Rescues Mitochondrial Dysfunction in a Murine Model*
**作者**: García, J. et al.
**摘要**: 通过腺相关病毒(AAV)递送重组TK2蛋白至TK2缺陷小鼠模型,显著改善线粒体DNA含量及肌肉功能,证明基因治疗的潜在可行性。
3. **文献名称**: *Structural Insights into Recombinant TK2 Mutants Associated with Mitochondrial DNA Depletion Syndrome*
**作者**: Johnson, R. et al.
**摘要**: 利用X射线晶体学解析TK2重组蛋白及其致病突变体的三维结构,揭示突变如何影响底物结合及酶活性,为药物设计提供依据。
4. **文献名称**: *In vitro Reconstitution of TK2-dependent Nucleotide Salvage Pathway Using Recombinant Proteins*
**作者**: Lee, S. et al.
**摘要**: 在体外重构了TK2重组蛋白与dGK(脱氧鸟苷激酶)协同作用的核苷酸补救通路,阐明其在维持线粒体dNTP池中的关键作用。
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**注意**:以上为示例文献,实际研究中请通过PubMed、Web of Science等平台检索真实文献。
**Background of Recombinant TK2 Protein**
Thymidine kinase 2 (TK2) is a mitochondrial enzyme encoded by the *TK2* gene, playing a critical role in the phosphorylation of thymidine and deoxycytidine—key steps in the salvage pathway of mitochondrial DNA (mtDNA) nucleotide synthesis. TK2 catalyzes the conversion of nucleosides to their monophosphate forms, ensuring a balanced supply of deoxyribonucleoside triphosphates (dNTPs) essential for mtDNA replication and repair. Defects in TK2 activity are linked to mitochondrial DNA depletion syndromes (MDS), a group of autosomal recessive disorders characterized by severe mtDNA copy number reduction, leading to progressive muscle weakness, encephalopathy, and organ failure.
Recombinant TK2 protein is produced using genetic engineering techniques, typically by expressing the human *TK2* gene in bacterial, insect, or mammalian cell systems. This approach allows large-scale production of the enzyme for functional studies, drug screening, and therapeutic exploration. Researchers utilize recombinant TK2 to investigate its kinetic properties, substrate specificity, and interactions with potential inhibitors or activators. Additionally, it serves as a tool to model TK2 deficiency in vitro, aiding in the development of targeted therapies, such as nucleoside bypass strategies to restore dNTP pools in MDS patients.
Recent advances in structural biology, including X-ray crystallography and cryo-EM, have leveraged recombinant TK2 to elucidate its 3D conformation and mechanism, providing insights into mutation-induced dysfunction. Efforts are also underway to explore enzyme replacement therapies (ERT) or gene therapies using recombinant TK2 to address mitochondrial disorders. Challenges remain in ensuring proper mitochondrial targeting and stability of the recombinant protein in vivo. Nonetheless, recombinant TK2 represents a pivotal resource for both understanding mitochondrial pathologies and advancing precision medicine for rare genetic diseases.
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