Recombinant HMGB1 with cytokine-stimulating activity

Journal of Immunological Methods
Jianhua LiHuan Yang

Abstract

We describe methods for the isolation, purification, and characterization of full-length high-mobility group box 1 (HMGB1) and truncated mutants expressed in bacteria and in mammalian Chinese Hamster Ovary (CHO) cells. HMGB1 is an abundant nuclear and cytoplasmic protein, highly conserved across species and widely distributed in eukaryotic cells from yeast to man. As a ubiquitous nuclear DNA binding protein, HMGB1 binds DNA, facilitates gene transcription, and stabilizes nucleosome structure. In addition to these intracellular roles, HMGB1 can be released into the extracellular milieu by activated innate immune cells (i.e., macrophages, monocytes) and functions as a mediator of lethal endotoxemia and sepsis. The proinflammatory cytokine activity of HMGB1 has become an intense area of research and recombinant protein can be a useful tool to probe HMGB1 functions. Due to its dipolar charged properties, HMGB1 isolated by some methods can be contaminated with bacterial products (such as CpG DNA or lipopolysaccharide [LPS]) that may interfere with immunological analyses. Here we report our newly developed methods for the isolation and purification of biologically active HMGB1 from bacteria or mammalian CHO cells that is essentially ...Continue Reading

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Citations

Jun 20, 2012·Neurological Sciences : Official Journal of the Italian Neurological Society and of the Italian Society of Clinical Neurophysiology·Wei LiChun-Hua Hang
May 28, 2013·Biochemical Pharmacology·Minghua YangDaolin Tang
Jun 16, 2010·Proceedings of the National Academy of Sciences of the United States of America·Huan YangKevin J Tracey
Jul 5, 2006·The Journal of Experimental Medicine·Shixin QinHuan Yang
Nov 14, 2012·Annals of the Rheumatic Diseases·Mei ZongHåkan Westerblad
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May 14, 2009·Zhong xi yi jie he xue bao = Journal of Chinese integrative medicine·Hui LiJian-hong Zhou
Jan 12, 2011·Annual Review of Immunology·Ulf Andersson, Kevin J Tracey
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