Construction and characterization of the HMGB1 mutant as a competitive antagonist to HMGB1 induced cytokines release.
Recent studies indicate that the High Mobility Group Box-1 protein (HMGB1) acts as a potent proinflammatory cytokine that contributes to the pathogenesis of diverse inflammatory and infectious disorders. The proinflammatory cytokine activity of HMGB1 has become a therapeutic target. In this study, we cloned the cDNA encoding human HMGB1 and constructed HMGB1 mutants using a one-step opposite direction PCR. The binding of the HMGB1 mutants to THP-1 cell and the cytokine activities of these HMGB1 mutants were observed. Results showed that the HMGB1 Mut (102-105), one of the HMGB1 mutants, in which amino acids 102-105 (FFLF) were replaced with two Glys, significantly decreased the full-length HMGB1 protein induced TNF-alpha release in human monocyte cultures. The results indicate that we have developed a novel recombinant HMGB1 mutant that competitively antagonizes the proinflammatory activity of HMGB1. This may be of significant importance in providing a new anti-inflammatory strategy for the treatment of severe sepsis and other inflammatory disorders.
Institute of Burn Research, Southwest Hospital, State Key Laboratory of Trauma, Burns and Combined Injury, Third Military Medical University, 30 Gaotanyan Street, Chongqing 400038, China. cqburn@yahoo.com.cn
Zhiqiang
Z
Yuan
Jing
J
Chen
Yan
Y
Zhang
Yizhi
Y
Peng
D000595:Amino Acid Sequence; D019943:Amino Acid Substitution; D000893:Anti-Inflammatory Agents / Q000494:pharmacology; D002460:Cell Line; D024243:HMGB1 Protein / Q000096:biosynthesis / Q000235:genetics* / Q000494:pharmacology*; D006801:Humans; D009000:Monocytes / Q000187:drug effects / Q000276:immunology; D009154:Mutation; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics* / Q000494:pharmacology*; D014409:Tumor Necrosis Factor-alpha / Q000037:antagonists & inhibitors* / Q000378:metabolism
D000893:Anti-Inflammatory Agents; D024243:HMGB1 Protein; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha