Screening HMGB1 promoter binding proteins in rat hepatic injury secondary  to severe acute pancreatitis using biotin-streptavidin system

作者：Yang L.; Zhang S.-R.; Wang H.-M.; Zhou X.-D.

来源：World Chinese Journal of Digestology, 2014, 22(25): 3796-3800.

DOI：10.11569/wcjd.v22.i25.3796

• Biotin-streptavidin
• High mobility group box 1 protein
• Promoter
• Severe acute pancreatitis

RESULTS: A total of 14 differential protein bands between the SAP and control groups were screened, 5 of which were identified as transcription related proteins by MS.AIM: To screen proteins that interact with high mobility group protein B1 (HMGB1) promoter in rat hepatic injury secondary to severe acute pancreatitis (SAP).METHODS: A rat model of SAP was generated by retrograde injection of 5 sodium taurocholate into the bilio-pancreatic duct. The SAP group and control group were executed simultaneously, and the liver nuclear extracts were prepared. PCR was used to amplify the biotin labeled tail probes of the HMGB1 promoter. The probes were incubated with cell nuclear extracts, and HMGB1 promoter-protein complexes were then separated using streptavidin conjugated magnetic beads. The proteins were eluted from probes with 0.25 mol/L and 1 mol/L NaCl, resolved using SDS-PAGE electrophoresis, and visualized by silver staining, and the differential bands were identified by mass spectrometry (MS).CONCLUSION: Proteins that interact with HMGB1 promoter in SAP-associated hepatic injury were acquired and identified, which have important value for the further study of regulatory mechanism of HMGB1.