Non-oxidizable chemokine-HMGB1, LPS-Free
HMGB1 is a nuclear protein that is released passively by necrotic cells, retained by apoptotic cells, and secreted actively by inflammatory cells (Dumitriu et al. HMGB1: guiding immunity from within. Trends Immunol 2005, 26: 381-7). HMGB1 is essential for life: Hmgb1 knockout mice die shortly after birth.
HMGB1 contains three conserved cysteine residues (C23, C45 and C106) and is a redox-sensitive (Yang et al, 2012). Chemotaxis-HMGB1 (the form with chemoattractanct activity) is completely reduced; cytokine-HMGB1 contains a disulfide bond between C23 and C45; further oxidation to sulfonates abrogate both activities.
In Non-oxidizable chemokine-HMGB1 all cysteines are replaced with serines: these replacements preserve chemoattractant activity in vitro and in vivo, eliminate the cytokine-inducing activity and make the protein resistant to inactivation by ROS (Venereau et al, 2012). The mutant protein is produced in E. coli.
It has the sequence:
[“MGKGDPKKPR GKMSSYAFFV QTSREEHKKK HPDASVNFSE FSKKSSERWK TMSAKEKGKF EDMAKADKAR YEREMKTYIP PKGETKKKFK DPNAPKRPPS AFFLFSSEYR PKIKGEHPGL SIGDVAKKLG EMWNNTAADD KQPYEKKAAK LKEKYEKDIA AYRAKGKPDA AKKGVVKAEK SKKKKEEEDD EEDEEDEEEE EEEEDEDEEE DDDDE”]
Molecular Mass: Non-oxidizable chemokine-HMGB1 consists of 215 amino acid residues and has a calculated molecular mass of approximately 24.8 kDa.
Structure: HMGB1 consists of two fairly rigid, L-shaped DNA-binding domains, each referred to as a ‘HMG box’, and an unstructured tail that ends with 30 consecutive negatively charged amino acids.
Purity: The purified protein is >95% homogeneous (electrophoresis and mass spectrometry). It contains no nucleic acids, nor IL-1beta.
Endotoxin Level: The purified protein is free from LPS (Cambrex Limulus Amoebocyte Assay QCL-1000, <0.4 ng LPS per mg protein).
Activity: Measured by its ability to induce migration. Maximal activity in the cell migration assay is obtained at 1 nM.
Buffer: Non-oxidizable chemokine-HMGB1 is lyophilized from 50 mM HEPES buffer, pH 7.9 and 500 mM NaCl.
Storage: 2-8°C. The protein once resuspended can be stored frozen (-20°C).This product is intended for research only, and cannot be used on humans.
This product is intended for research only, and cannot be used on humans.
Publications where this product was used:
Venereau et al (2012) Mutually exclusive redox forms of HMGB1 promote cell recruitment or proinflammatory cytokine release. Journal of Experimental Medicine (2012).
Tirone et al (2017) High mobility group box 1 orchestrates tissue regeneration via CXCR4. J.Exp Med
Download Non-oxidizable chemokine-HMGB1 Datasheet
Complete Name: High Mobility Group 1 Mutant
Other Names: 3S, Amphoterin-1, High mobility group protein 1 mutant ,3S Mutant, Triple Serine Mutant, HMG-1, SBP-1, high-mobility group box 1, Amphoterin, HMG1, recombinant HMGB1