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.
Yuan
Zhiqiang
Z
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
Chen
Jing
J
Zhang
Yan
Y
Peng
Yizhi
Y
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