Recombinant HMGB1 with cytokine-stimulating activity.

Published

Journal Article

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 free of contaminants. This strategy provides an important advance in methodology to facilitate future HMGB1 studies.

Full Text

Duke Authors

Cited Authors

  • Li, J; Wang, H; Mason, JM; Levine, J; Yu, M; Ulloa, L; Czura, CJ; Tracey, KJ; Yang, H

Published Date

  • June 2004

Published In

Volume / Issue

  • 289 / 1-2

Start / End Page

  • 211 - 223

PubMed ID

  • 15251426

Pubmed Central ID

  • 15251426

International Standard Serial Number (ISSN)

  • 0022-1759

Digital Object Identifier (DOI)

  • 10.1016/j.jim.2004.04.019

Language

  • eng

Conference Location

  • Netherlands