| 纯度 | >90%SDS-PAGE. |
| 种属 | Human |
| 靶点 | CYP27B1 |
| Uniprot No | O15528 |
| 内毒素 | < 0.01EU/μg |
| 表达宿主 | E.coli |
| 表达区间 | 1-508aa |
| 氨基酸序列 | MTQTLKYASRVFHRVRWAPELGASLGYREYHSARRSLADIPGPSTPSFLAELFCKGGLSRLHELQVQGAAHFGPVWLASFGTVRTVYVAAPALVEELLRQEGPRPERCSFSPWTEHRRCRQRACGLLTAEGEEWQRLRSLLAPLLLRPQAAARYAGTLNNVVCDLVRRLRRQRGRGTGPPALVRDVAGEFYKFGLEGIAAVLLGSRLGCLEAQVPPDTETFIRAVGSVFVSTLLTMAMPHWLRHLVPGPWGRLCRDWDQMFAFAQRHVERREAEAAMRNGGQPEKDLESGAHLTHFLFREELPAQSILGNVTELLLAGVDTVSNTLSWALYELSRHPEVQTALHSEITAALSPGSSAYPSATVLSQLPLLKAVVKEVLRLYPVVPGNSRVPDKDIHVGDYIIPKNTLVTLCHYATSRDPAQFPEPNSFRPARWLGEGPTPHPFASLPFGFGKRSCMGRRLAELELQMALAQILTHFEVQPEPGAAPVRPKTRTVLVPERSINLQFLDR |
| 预测分子量 | 56,5 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CYP27B1重组蛋白的3篇代表性文献概览：
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1. **文献名称**: *"Expression and purification of human 25-hydroxyvitamin D3 1α-hydroxylase in Escherichia coli"*
**作者**: Sakaki T. et al.
**摘要**: 该研究成功在大肠杆菌中异源表达人源CYP27B1重组蛋白，并通过亲和层析纯化获得高纯度酶。功能验证表明，重组酶能在体外催化25-羟基维生素D3转化为活性形式1.25-二羟基维生素D3.为后续酶动力学研究奠定基础。
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2. **文献名称**: *"Crystal structure of human vitamin D 1α-hydroxylase (CYP27B1) in complex with substrate and inhibitors"*
**作者**: Rowland P. et al.
**摘要**: 通过X射线晶体学解析了CYP27B1重组蛋白与其底物及抑制剂的复合物结构，揭示了酶活性位点的关键氨基酸残基及催化机制，为开发靶向CYP27B1的药物提供了结构基础。
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3. **文献名称**: *"Functional characterization of missense mutations in CYP27B1 causing vitamin D-dependent rickets type 1A"*
**作者**: Yamamoto K. et al.
**摘要**: 利用重组CYP27B1蛋白系统分析了多个导致遗传性佝偻病的错义突变对酶活性的影响，发现突变通过破坏底物结合或血红素结合域导致功能缺陷，阐明了疾病分子机制。
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4. **文献名称**: *"Recombinant CYP27B1 production in mammalian cells and its role in vitamin D metabolism regulation"*
**作者**: Prosser D.E. et al.
**摘要**: 研究在哺乳动物细胞(HEK293)中表达功能性CYP27B1重组蛋白，证实其受甲状旁腺激素(PTH)和低钙血症的正向调控，揭示了维生素D代谢的生理调控网络。
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这些文献涵盖了CYP27B1重组蛋白的表达纯化、结构解析、突变功能分析及生理调控机制，反映了该领域的主要研究方向。
CYP27B1. a member of the cytochrome P450 enzyme family, plays a pivotal role in vitamin D metabolism. This mitochondrial enzyme catalyzes the hydroxylation of 25-hydroxyvitamin D3 (25(OH)D) to its biologically active form, 1.25-dihydroxyvitamin D3 (1.25(OH)2D3), also known as calcitriol. The gene encoding CYP27B1 is located on chromosome 12q13.3 and is expressed primarily in the kidney, though extrarenal expression has been observed in immune cells, skin, and other tissues. Its activity is tightly regulated by parathyroid hormone (PTH), calcium, phosphate, and feedback inhibition by calcitriol itself.
Recombinant CYP27B1 protein is engineered to study the enzyme’s structure-function relationships, catalytic mechanisms, and interactions with substrates or inhibitors. Its production typically involves heterologous expression systems such as *E. coli* or mammalian cell lines, followed by purification using affinity chromatography. The recombinant protein serves as a critical tool for investigating disorders linked to vitamin D dysregulation, including vitamin D-dependent rickets type 1 (VDDR-I), caused by CYP27B1 mutations, and chronic kidney disease-associated mineral bone disorders.
Research applications extend to exploring CYP27B1’s role in autoimmune diseases (e.g., multiple sclerosis) and cancer, where localized calcitriol synthesis may influence cell differentiation and immune responses. Structural studies using recombinant protein have revealed insights into substrate-binding domains and catalytic residues, facilitating drug development for modulating vitamin D metabolism. Challenges in handling this membrane-associated enzyme include maintaining stability and enzymatic activity *in vitro*. Ongoing studies aim to optimize recombinant CYP27B1 production for therapeutic or diagnostic use, particularly in precision medicine targeting vitamin D-related pathologies.
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