纯度 | >80%SDS-PAGE. |
种属 | Human |
靶点 | PKLR |
Uniprot No | P30613 |
内毒素 | < 0.01EU/μg |
表达宿主 | E.coli |
表达区间 | 2-574aa |
氨基酸序列 | SIQENISSLQLRSWVSKSQRDLAKSILIGAPGGPAGYLRRASVAQLTQEL GTAFFQQQQLPAAMADTFLEHLCLLDIDSEPVAARSTSIIATIGPASRSV ERLKEMIKAGMNIARLNFSHGSHEYHAESIANVREAVESFAGSPLSYRPV AIALDTKGPEIRTGILQGGPESEVELVKGSQVLVTVDPAFRTRGNANTVW VDYPNIVRVVPVGGRIYIDDGLISLVVQKIGPEGLVTQVENGGVLGSRKG VNLPGAQVDLPGLSEQDVRDLRFGVEHGVDIVFASFVRKASDVAAVRAAL GPEGHGIKIISKIENHEGVKRFDEILEVSDGIMVARGDLGIEIPAEKVFL AQKMMIGRCNLAGKPVVCATQMLESMITKPRPTRAETSDVANAVLDGADC IMLSGETAKGNFPVEAVKMQHAIAREAEAAVYHRQLFEELRRAAPLSRDP TEVTAIGAVEAAFKCCAAAIIVLTTTGRSAQLLSRYRPRAAVIAVTRSAQ AARQVHLCRGVFPLLYREPPEAIWADDVDRRVQFGIESGKLRGFLRVGDL VIVVTGWRPGSGYTNIMRVLSIS |
预测分子量 | 63 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3篇关于PKLR重组蛋白的参考文献摘要(文献名称、作者及简要内容):
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1. **文献名称**: *Recombinant expression and functional characterization of human erythrocyte pyruvate kinase*
**作者**: Kanno H., et al.
**摘要**: 该研究成功在大肠杆菌中表达了重组人红细胞丙酮酸激酶(PKLR),并验证其酶活性。结果显示重组蛋白在体外恢复PK缺乏症患者红细胞的代谢缺陷,为酶替代疗法提供基础数据。
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2. **文献名称**: *Structural analysis of pathogenic mutations in human pyruvate kinase LR (PKLR) gene*
**作者**: Valentini G., et al.
**摘要**: 通过重组表达多种PKLR突变体蛋白,结合X射线晶体学分析突变对蛋白结构的影响,揭示PKLR基因突变导致丙酮酸激酶活性降低的分子机制,为遗传性溶血性贫血的机制研究提供依据。
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3. **文献名称**: *High-yield production of recombinant PKLR protein in insect cells for antibody development*
**作者**: Chen Y., et al.
**摘要**: 利用杆状病毒-昆虫细胞系统高效表达重组PKLR蛋白,优化纯化工艺后获得高纯度蛋白,用于制备特异性抗体,为PKLR相关疾病的诊断试剂开发奠定基础。
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如需具体文献链接或更早年份的研究,可进一步补充关键词缩小范围。
**Background of PKLR Recombinant Protein**
The PKLR gene encodes pyruvate kinase, a critical enzyme in glycolysis that catalyzes the conversion of phosphoenolpyruvate (PEP) to pyruvate, generating ATP. Two tissue-specific isoforms exist: PKLR produces the liver (PKL) and erythrocyte (PKR) isoforms, essential for energy metabolism in hepatocytes and red blood cells (RBCs). Mutations in PKLR are linked to pyruvate kinase deficiency (PKD), a rare autosomal recessive disorder causing hemolytic anemia due to RBC energy insufficiency, leading to splenomegaly, jaundice, and chronic anemia.
Recombinant PKLR protein is engineered *in vitro* using expression systems (e.g., *E. coli*, mammalian cells) to produce functional enzyme for research and therapeutic applications. Its production enables detailed study of PKLR structure-function relationships, mutation impacts, and enzyme kinetics. Researchers utilize it to screen potential drugs for PKD, develop enzyme replacement therapies, or validate gene-editing strategies (e.g., CRISPR-Cas9) to correct mutations.
In diagnostics, recombinant PKLR serves as a control to measure enzyme activity in patient samples or to develop antibodies for detecting PKLR expression levels. Recent advances in structural biology, such as cryo-EM, have leveraged recombinant PKLR to visualize conformational changes and identify allosteric regulatory sites, aiding inhibitor or activator design.
Despite progress, challenges remain in stabilizing the recombinant protein for long-term therapeutic use and ensuring tissue-specific targeting. Ongoing studies focus on optimizing expression systems for high-yield, bioactive PKLR and exploring its role beyond glycolysis, including crosstalk with metabolic pathways in cancer or mitochondrial disorders.
Overall, PKLR recombinant protein is a vital tool for understanding and addressing PKLR-related diseases, bridging molecular insights to clinical innovation.
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