Fully reduced 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.
HMGB1 is essential for life: Hmgb1 knockout mice die shortly after Birth (Dumitriu et al. HMGB1: guiding immunity from within. Trends Immunol 2005, 26: 381-7).
The two different activities of HMGB1 as a chemoattractanct of motile cells and as inducer of cytokines have been attributed to different biochemical forms. Fully reduced HMGB1 complete notation: HMGB1C23hC45hC106h – Antoine J.,2014) is completely reduced and has chemoattractanct activity; Disulfide HMGB1 contains a disulfide bond between C23 and C45 (complete notation: HMGB1C23-C45C106h – Antoine J.,2014) and can induce cytokine and chemokine production in monocytes and other inflammatory cells.
Fully reduced HMGB1 had previously been sold by HMGBiotech as Chemotaxis-HMGB1 LPS free and tested in migration assay, but is otherwise the same identical product.
Fully reduced HMGB1 behaves as a trigger of inflammation, attracting inflammatory cells, and of tissue repair, attracting stem cells and promoting their proliferation. Moreover, Fully reduced HMGB1 activates dedritic cells and promotes their functional maturation and migration to lymph nodes.
However, and contrary to early reports, pure Fully reduced HMGB1 does not promote cytokine secretion. Cytokine secretion is only promoted by disulfide HMGB1 (the form containing disulfide bond).
The Fully reduced HMGB1 we provide is the natural protein, with no tags or additional amino acids. It is produced in E.coli from an expression plasmid coding for the human protein.
It has the sequence:
[“MGKGDPKKPR GKMSSYAFFV QTCREEHKKK HPDASVNFSE FSKKCSERWK TMSAKEKGKF EDMAKADKAR YEREMKTYIP PKGETKKKFK DPNAPKRPPS AFFLFCSEYR PKIKGEHPGL SIGDVAKKLG EMWNNTAADD KQPYEKKAAK LKEKYEKDIA AYRAKGKPDA AKKGVVKAEK SKKKKEEEDD EEDEEDEEEE EEEEDEDEEE DDDDE” ]
Molecular Mass: HMGB1 consists of 215 amino acid residues and has a calculated molecular mass of approximately 24.8 kDa. It migrates at a position of approximately 30 kD in SDS-PAGE gels, possibly because of the unusual number of positively charged amino acids it contains.
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.
Species specificity: the sequence and structure of HMGB1 have been strictly conserved during evolution, and HMGB1 proteins from all mammals are almost identical (Sessa L, Bianchi ME. The evolution of High Mobility Group Box (HMGB) chromatin proteins in multicellular animals. Gene 2007, 387:133-40).
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, or 25 ng/ml (Palumbo et al. Extracellular HMGB1, a signal of tissue damage, induces mesoangioblast migration and proliferation. J Cell Biol 2004, 164: 441-9).
Buffer: 50 mM HEPES-Na pH 7.9, 500 mM NaCl, 0.5 mM DTT.
Storage: the protein is shipped lyophilized. Once resuspended can be stored frozen at -20°C. To avoid cysteine oxidation, DTT 0.5mM is added during protein purification.
This product is intended for research only, and cannot be used on humans.
Publications where this product was used:
16th June 2023
HMGB1●CXCL12 : the first fuzzy chemokines heterocomplex reported so far
HMGBiotech Srl participated in a recent study in which, through an integrative structural approach,
molecular details of HMGB1●CXCL12 heterocomplex formation were unveiled.
This dynamic complex is formed equimolarly, contrary to previous assumptions.
Structured and unstructured HMGB1 regions interact with the dimerization surface of CXCL12. This work elucidated how the acidic IDR is involved in HMGB1●CXCL12 complex formation.The findings suggest that interfering with the interactions in the HMGB1●CXCL12 complex could potentially inhibit its detrimental effects in inflammatory conditions. Read the full article about the study…
Limana et al. Exogenous High-Mobility Group Box 1 protein induces myocardial regeneration following infarction via enhanced cardiac c-kit cell proliferation and differentiation. Circ Res 2005, 97:73-83.
Other publications: 1 , 2 , 3 , 4 , 5 , 6 , 7 , 8 , 9 , 10, 11 , 12
Complete Name: High Mobility Group 1
Other Names: Amphoterin-1, High mobility group protein 1,HMG-1, SBP-1, high mobility group protein B1, high-mobility group box 1, Amphoterin, HMG1, HMGB1, SBP-1, recombinant HMGB1