Expression and purification of functional HMGB1 A box by fusion with SUMO

Ge,Wen‑Song; Fan,Jian‑Gao; Chen,Ying‑Wei; Xu,Lei‑Ming

doi:10.3892/mmr.2015.4308

Expression and purification of functional HMGB1 A box by fusion with SUMO

Authors:
- Wen‑Song Ge
- Jian‑Gao Fan
- Ying‑Wei Chen
- Lei‑Ming Xu
View Affiliations

Affiliations: Department of Gastroenterology, Shanghai Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200092, P.R. China
Published online on: September 9, 2015 https://doi.org/10.3892/mmr.2015.4308
Pages: 6527-6532
Copyright: © Ge et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

High‑mobility‑group‑box chromosomal protein 1 (HMGB1) is a ubiquitous and abundant nuclear protein in eukaryotic cells. Nuclear HMGB1 serves an important role in maintaining nuclear stability under stress. However, extracellular HMGB1 exerts actions, which are distinctly different compared with these intracellular functions. HMGB1, when released extracellularly, is a potent innate signal, which initiates host defense mechanisms or tissue regeneration. HMGB1 has two DNA‑binding domains: HMG A box and B box. The HMGB1 A box exhibits an antagonistic, anti‑inflammatory effect, and is a potential therapeutic target, however, the large‑scale expression and purification of the HMGB1 A box with high efficiency remains to be reported. In the present study, a SUMO‑fusion expression system was used to express and purify high levels of functional HMGB1 A box to meet the requirements of therapeutic protein production.

View Figures

View References

1	Bustin M, Lehn DA and Landsman D: Structural features of the HMG chromosomal proteins and their genes. Biochim Biophys Acta. 1049:231–243. 1990. View Article : Google Scholar : PubMed/NCBI
2	Boonyaratanakornkit V, Melvin V, Prendergast P, Altmann M, Ronfani L, Bianchi ME, Taraseviciene L, Nordeen SK, Allegretto EA and Edwards DP: High-mobility group chromatin proteins 1 and 2 functionally interact with steroid hormone receptors to enhance their DNA binding in vitro and transcriptional activity in mammalian cells. Mol Cell Biol. 18:4471–4487. 1998. View Article : Google Scholar : PubMed/NCBI
3	Melvin VS and Edwards DP: Coregulatory proteins in steroid hormone receptor action: The role of chromatin high mobility group proteins HMG-1 and -2. Steroids. 64:576–586. 1999. View Article : Google Scholar : PubMed/NCBI
4	Vaccari T, Beltrame M, Ferrari S and Bianchi ME: Hmg4, a new member of the Hmg1/2 gene family. Genomics. 49:247–252. 1998. View Article : Google Scholar : PubMed/NCBI
5	Funayama A, Shishido T, Netsu S, Narumi T, Kadowaki S, Takahashi H, Miyamoto T, Watanabe T, Woo CH, Abe J, et al: Cardiac nuclear high mobility group box 1 prevents the development of cardiac hypertrophy and heart failure. Cardiovasc Res. 99:657–664. 2013. View Article : Google Scholar : PubMed/NCBI
6	Tsung A, Tohme S and Billiar TR: High mobility group box-1 in sterile inflammation. J Intern Med. 276:425–443. 2014. View Article : Google Scholar : PubMed/NCBI
7	Dumitriu IE, Baruah P, Manfredi AA, Bianchi ME and Rovere-Querini P: HMGB1: Guiding immunity from within. Trends Immunol. 26:381–387. 2005. View Article : Google Scholar : PubMed/NCBI
8	Yang D, Chen Q, Yang H, Tracey KJ, Bustin M and Oppenheim JJ: High mobility group box-1 protein induces the migration and activation of human dendritic cells and acts as an alarmin. J Leukoc Biol. 81:59–66. 2007. View Article : Google Scholar
9	Yang H, Wang H, Czura CJ and Tracey KJ: HMGB1 as a cytokine and therapeutic target. J Endotoxin Res. 8:469–472. 2002. View Article : Google Scholar
10	Andersson U and Tracey KJ: HMGB1 in sepsis. Scand J Infect Dis. 35:577–584. 2003. View Article : Google Scholar : PubMed/NCBI
11	Kokkola R, Li J, Sundberg E, Aveberger AC, Palmblad K, Yang H, Tracey KJ, Andersson U and Harris HE: Successful treatment of collagen-induced arthritis in mice and rats by targeting extracellular high mobility group box chromosomal protein 1 activity. Arthritis Rheum. 48:2052–2058. 2003. View Article : Google Scholar : PubMed/NCBI
12	Wang H, Bloom O, Zhang M, Vishnubhakat JM, Ombrellino M, Che J, Frazier A, Yang H, Ivanova S, Borovikova L, et al: HMG-1 as a late mediator of endotoxin lethality in mice. Science. 285:248–251. 1999. View Article : Google Scholar : PubMed/NCBI
13	Huang Y, Yin H, Han J, Huang B, Xu J, Zheng F, Tan Z, Fang M, Rui L, Chen D, et al: Extracellular hmgb1 functions as an innate immune-mediator implicated in murine cardiac allograft acute rejection. Am J Transplant. 7:799–808. 2007. View Article : Google Scholar : PubMed/NCBI
14	Andrassy M, Volz HC, Igwe JC, Funke B, Eichberger SN, Kaya Z, Buss S, Autschbach F, Pleger ST, Lukic IK, et al: High-mobility group box-1 in ischemia-reperfusion injury of the heart. Circulation. 117:3216–3226. 2008. View Article : Google Scholar : PubMed/NCBI
15	Andersson U and Tracey KJ: HMGB1 is a therapeutic target for sterile inflammation and infection. Annu Rev Immunol. 29:139–162. 2011. View Article : Google Scholar : PubMed/NCBI
16	Yang H, Ochani M, Li J, Qiang X, Tanovic M, Harris HE, Susarla SM, Ulloa L, Wang H, DiRaimo R, et al: Reversing established sepsis with antagonists of endogenous high-mobility group box 1. Proc Natl Acad Sci USA. 101:296–301. 2004. View Article : Google Scholar :
17	Butt TR, Edavettal SC, Hall JP and Mattern MR: SUMO fusion technology for difficult-to-express proteins. Protein Expr Purif. 43:1–9. 2005. View Article : Google Scholar : PubMed/NCBI
18	Johnson ES: Protein modification by SUMO. Annu Rev Biochem. 73:355–382. 2004. View Article : Google Scholar : PubMed/NCBI
19	Hochstrasser M: Evolution and function of ubiquitin-like protein-conjugation systems. Nat Cell Biol. 2:E153–E157. 2000. View Article : Google Scholar : PubMed/NCBI
20	Jentsch S and Pyrowolakis G: Ubiquitin and its kin: How close are the family ties? Trends Cell Biol. 10:335–342. 2000. View Article : Google Scholar : PubMed/NCBI
21	Müller S, Hoege C, Pyrowolakis G and Jentsch S: SUMO, ubiquitin's mysterious cousin. Nat Rev Mol Cell Biol. 2:202–210. 2001. View Article : Google Scholar : PubMed/NCBI
22	Malakhov MP, Mattern MR, Malakhova OA, Drinker M, Weeks SD and Butt TR: SUMO fusions and SUMO-specific protease for efficient expression and purification of proteins. J Struct Funct Genomics. 5:75–86. 2004. View Article : Google Scholar : PubMed/NCBI
23	Zuo X, Mattern MR, Tan R, Li S, Hall J, Sterner DE, Shoo J, Tran H, Lim P, Sarafianos SG, et al: Expression and purification of SARS coronavirus proteins using SUMO-fusions. Protein Expr Purif. 42:100–110. 2005. View Article : Google Scholar : PubMed/NCBI
24	Zhang Z, Zhang L, Zhou C and Wu H: Ketamine inhibits LPS-induced HGMB1 release in vitro and in vivo. Int Immunopharmacol. 23:14–26. 2014. View Article : Google Scholar : PubMed/NCBI
25	Kong X, Yuan H, Wu X, Zhang J, Zhou H, Wang M, Liu Y and Jin X: High-mobility-group box protein 1A box reduces development of sodium laurate-induced thromboangiitis obliterans in rats. J Vasc Surg. 57:194–204. 2013. View Article : Google Scholar