Study Of HMGN2 Subcellular Distribution And Its Recombinant Immunotoxin

Endogeneous antimicrobial peptides are important mediators in innate immunity. Finding new antimicrobial peptides is most scientists'hope. The high Mobility Group Chromosal protein N2 (HMGN2) protein is one of the most abundant and well characterized class of nonhistone nuclear proteins which seem to function as architectural elements in chromatin, is found in all of the cells of higher eukaryotes . However, the biological role of this protein has not been fully defined. Recently Our group found that the HMGN2 had antimicrobial activity through isolation and characterization of antimicrobial peptides from human LAK cell and human cervical mucus. To confirm HMGN2 taking part in innate immunity as an antimicrobial peptides, we studied the subcellular localization of HMGN2 in human monocytes and Hela cells. Using the tumor-targeting capacity of HMGN2 α-helical domain, we also constructed a recombinant immunotoxin and examined its cytotoxicity to cancer cells. This study includes three parts.Part I is to prepare HMGN2 polyclonal antibodies and determine the subcellular localization of HMGN2 in human monocytes. The recombinant prokaryotic expression vector pGEX-1λT-HMGN2 was constructed and E.coli-based product of GST-HMGN2 fusion protein was prepared and used to immunize rabbit for producing the anti-serum against HMGN2. The polyclonal antibodies were partially purified by caprylic acid and ammonium sulfate precipitation. The titter of specific polyclonal antibodies against HMGN2 was detected by ELISA. The immunocytochemical staining was performed to determine the distribution of HMGN2 in THP-1 cells. Gel...