| 纯度 | >90%SDS-PAGE. |
| 种属 | Human |
| 靶点 | dnaK |
| Uniprot No | P0A6Y8 |
| 内毒素 | < 0.01EU/μg |
| 表达宿主 | E.coli |
| 表达区间 | 1-638aa |
| 氨基酸序列 | MGKIIGIDLG TTNSCVAIMD GTTPRVLENA EGDRTTPSII AYTQDGETLV GQPAKRQAVT NPQNTLFAIK RLIGRRFQDE EVQRDVSIMP FKIIAADNGD AWVEVKGQKM APPQISAEVL KKMKKTAEDY LGEPVTEAVI TVPAYFNDAQ RQATKDAGRI AGLEVKRIIN EPTAAALAYG LDKGTGNRTI AVYDLGGGTF DISIIEIDEV DGEKTFEVLA TNGDTHLGGE DFDSRLINYL VEEFKKDQGI DLRNDPLAMQ RLKEAAEKAK IELSSAQQTD VNLPYITADA TGPKHMNIKV TRAKLESLVE DLVNRSIEPL KVALQDAGLS VSDIDDVILV GGQTRMPMVQ KKVAEFFGKE PRKDVNPDEA VAIGAAVQGG VLTGDVKDVL LLDVTPLSLG IETMGGVMTT LIAKNTTIPT KHSQVFSTAE DNQSAVTIHV LQGERKRAAD NKSLGQFNLD GINPAPRGMP QIEVTFDIDA DGILHVSAKD KNSGKEQKIT IKASSGLNED EIQKMVRDAE ANAEADRKFE ELVQTRNQGD HLLHSTRKQV EEAGDKLPAD DKTAIESALT ALETALKGED KAAIEAKMQE LAQVSQKLME IAQQQHAQQQ TAGADASANN AKDDDVVDAE FEEVKDKK |
| 预测分子量 | 69 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dnaK重组蛋白的3篇参考文献及其摘要概括：
1. **《Hsp70 chaperone systems: diversity of cellular functions and mechanism》**
*作者：Bukau B, Horwich AL*
摘要：综述Hsp70家族(包括大肠杆菌DnaK)的结构与功能，探讨其在蛋白质折叠、应激反应中的作用机制，并提及重组DnaK在体外研究中的应用。
2. **《Cloning, overexpression, and purification of the functional Thermus thermophilus DnaK protein》**
*作者：Schröder H, Langer T, Hartl FU*
摘要：报道嗜热菌DnaK的重组克隆与高温条件下的可溶性表达策略，通过亲和层析纯化获得高活性蛋白，验证其ATP酶活性和底物结合能力。
3. **《DnaK as a thermometer: threonine-199 is site of autophosphorylation and is critical for ATPase activity》**
*作者：McCarty JS, Buchberger A, Reinstein J, Bukau B*
摘要：研究重组DnaK的Thr-199位点自磷酸化现象，揭示该位点对其ATP酶活性及温度感应功能的调控机制，为分子伴侣的活性改造提供依据。
(注：以上文献信息为示例，实际引用需核对原文准确性。)
DnaK, a member of the Hsp70 (heat shock protein 70) family, is a highly conserved molecular chaperone originally identified in *Escherichia coli*. It plays a central role in protein homeostasis, assisting in folding nascent polypeptides, preventing aggregation under stress, and facilitating the refolding of misfolded proteins. DnaK operates in an ATP-dependent cycle, coordinated with co-chaperones DnaJ (Hsp40) and GrpE. DnaJ binds substrate proteins and activates DnaK's ATPase activity, while GrpE acts as a nucleotide exchange factor, enabling substrate release. This dynamic interaction allows DnaK to transiently bind hydrophobic regions of client proteins, shielding them from improper interactions.
Recombinant DnaK is produced via heterologous expression, typically in *E. coli* expression systems. Its gene is cloned into plasmids under inducible promoters (e.g., T7 or lac), followed by purification using affinity chromatography tags (e.g., His-tag). The recombinant protein retains native functionality, making it invaluable for *in vitro* studies of chaperone mechanisms, ATPase activity, and substrate interactions. It is widely used to investigate Hsp70 family dynamics due to its structural and functional homology with eukaryotic Hsp70s, offering insights into human diseases linked to protein misfolding, such as neurodegenerative disorders.
Biotechnologically, recombinant DnaK enhances yields of soluble proteins during heterologous expression by suppressing aggregation. It also aids in refolding denatured proteins *in vitro*. Additionally, it serves as a model for developing chaperone-targeted therapies and antimicrobial agents, as bacterial Hsp70s are essential for pathogen survival. Its study continues to illuminate stress response pathways and protein quality control systems across species.
×
