CKB Human
Description
Introduction and Overview
Creatine Kinase B (CKB), also known as Brain-type Creatine Kinase, is a cytoplasmic enzyme critical for cellular energy homeostasis. In humans, it is encoded by the CKB gene (OMIM: 123280) and exists as a homodimer in brain, smooth muscle, and other tissues, while forming heterodimers with muscle-type creatine kinase (CKM) in the heart . Recombinant CKB Human is produced in E. coli as a non-glycosylated protein, with two primary isoforms:
| Parameter | PKA-089 | CKI-274 |
|---|---|---|
| Molecular Mass | 42.6 kDa | 44.8 kDa |
| His-Tag | No | Yes (20 residues) |
| Purity | >90% (SDS-PAGE) | >95% (SDS-PAGE) |
| Storage | -20°C (long-term), 4°C (short-term) | -18°C |
CKB reversibly catalyzes the transfer of phosphate between ATP and creatine phosphate, maintaining ATP/ADP ratios in energy-demanding tissues .
Crystallographic Insights
CKB’s crystal structures reveal dynamic conformations:
The His-tagged variant (CKI-274) has been co-crystallized with a covalent inhibitor (KLU), revealing interactions critical for drug design .
Energy Metabolism
CKB regulates mitochondrial ATP production by suppressing AKT activation, reducing mitochondrial calcium (mCa²⁺) levels, and inhibiting mitochondrial permeability transition pore (mPTP) activity . This mechanism sustains ATP synthesis in cells reliant on oxidative phosphorylation (e.g., neurons, cancer cells) .
Cancer Progression
CKB is implicated in tumor growth and metastasis:
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Osteosarcoma: Overexpression promotes proliferation, invasion, and apoptosis resistance via p53 pathway suppression .
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Breast Cancer: HIF-1α-dependent CKB expression enhances metastasis by upregulating glycolysis and oxidative phosphorylation .
Research Applications and Experimental Models
CKB Human is used in:
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Cell Culture Assays:
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In Vivo Models:
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Knockdown/Overexpression: Silencing CKB reduces CK activity and sensitizes cells to cyclocreatine (cCr) .
Targeting CKB in Cancer
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Cyclocreatine (cCr): Inhibits CK activity, reducing tumor growth and metastasis in breast cancer models . Synergy with doxorubicin observed in triple-negative breast cancer (TNBC) .
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Mitochondrial ATP Modulation: CKB suppression may enhance chemotherapy efficacy by impairing energy metabolism in cancer cells .
Aging and Metabolic Disorders
Low CKB expression in aging correlates with mitochondrial dysfunction, suggesting potential therapeutic targets for age-related diseases .
Production and Purification
Recombinant CKB Human is purified via chromatography and formulated in Tris-HCl buffer with glycerol and DTT . Stability is maintained at -20°C, with freeze-thaw cycles avoided .
| Formulation Component | Concentration | Purpose |
|---|---|---|
| Tris-HCl Buffer | 20 mM (pH 8.0) | Maintain pH stability |
| Glycerol | 10% | Protein stabilization |
| DTT | 1 mM | Reduce oxidative stress |
Product Specs
Product Science Overview
Creatine kinase (CK) is an enzyme that plays a crucial role in cellular energy homeostasis. It catalyzes the reversible conversion of creatine and adenosine triphosphate (ATP) to phosphocreatine and adenosine diphosphate (ADP). This reaction is vital for the rapid regeneration of ATP, which is essential for various cellular processes, including muscle contraction and intracellular energy transport.
There are three main isoforms of creatine kinase, each predominantly found in different tissues:
- CK-MM: Found mainly in skeletal muscle.
- CK-MB: Found primarily in cardiac muscle.
- CK-BB: Found mainly in the brain and smooth muscle.
CK-BB is the isoform of creatine kinase that is predominantly expressed in the brain. It plays a significant role in maintaining the energy balance within brain cells, particularly neurons. The high energy demands of the brain make CK-BB essential for proper brain function.
Recombinant human creatine kinase brain (CK-BB) is a form of the enzyme that is produced using recombinant DNA technology. This involves inserting the gene encoding CK-BB into a host organism, such as Escherichia coli (E. coli), which then expresses the protein. The recombinant protein is subsequently purified for use in research and clinical applications.
The production of recombinant CK-BB typically involves the following steps:
- Gene Cloning: The gene encoding human CK-BB is cloned into an expression vector.
- Transformation: The expression vector is introduced into a host organism, such as E. coli.
- Expression: The host organism is cultured under conditions that induce the expression of CK-BB.
- Purification: The expressed CK-BB protein is purified using techniques such as affinity chromatography.
Recombinant CK-BB has several important applications:
- Research: It is used in studies investigating the role of CK-BB in brain metabolism and neurodegenerative diseases.
- Clinical Diagnostics: CK-BB levels can be measured in clinical settings to assess brain damage or disease.
- Drug Development: It is used in the development of drugs targeting energy metabolism in the brain.
