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Native versus recombinant high-mobility group B1 proteins: functional activity in vitro.
Klaus Zimmermann1, Dirk Völkel, Sabrina Pable
1Baxter BioScience, Industriestrasse 72, 1220 Vienna, Austria.
Inflammation
|January 28, 2005
Summary
Native and recombinant High Mobility Group Box 1 (HMGB1) proteins both stimulate tumor necrosis factor-alpha release and cell proliferation. Recombinant HMGB1s showed greater efficacy in these functional assays.
Area of Science:
- Biochemistry
- Immunology
- Cell Biology
Background:
- High Mobility Group Box 1 (HMGB1) is a nuclear protein with extracellular functions.
- Understanding the functional activity of HMGB1 from different sources is crucial for its therapeutic applications.
Purpose of the Study:
- To compare the functional activity of native eukaryotic HMGB1 with prokaryotic recombinant HMGB1.
- To investigate the in vitro biological effects of HMGB1 on immune and somatic cells.
Main Methods:
- Cloning and bacterial expression of human and murine HMGB1 cDNAs.
- Purification of native HMGB1 from calf thymus and Chinese hamster ovary (CHO) cells.
- In vitro assays measuring tumor necrosis factor-alpha (TNF-alpha) release in human whole blood and THP-1 cells, and NIH/3T3 cell proliferation.
Main Results:
- Both native and recombinant HMGB1 proteins dose-dependently induced TNF-alpha release in human blood and THP-1 cells.
- Recombinant HMGB1 proteins demonstrated higher potency in inducing TNF-alpha release compared to native HMGB1.
- Native and recombinant HMGB1s stimulated NIH/3T3 cell proliferation.
Conclusions:
- Native eukaryotic HMGB1 proteins exhibit similar biological activities in vitro as recombinant prokaryotic HMGB1 proteins.
- Recombinant HMGB1s are more potent in inducing TNF-alpha release and cell proliferation.
- These findings highlight the conserved functional properties of HMGB1 across different sources.