期刊
INSECT BIOCHEMISTRY AND MOLECULAR BIOLOGY
卷 43, 期 7, 页码 612-625出版社
PERGAMON-ELSEVIER SCIENCE LTD
DOI: 10.1016/j.ibmb.2013.03.013
关键词
Gloverin; Lipopolysaccharide; Circular dichroism; Antibacterial; Random coil; alpha-helix; Bombyx mori
资金
- National Institutes of Health [GM066356]
- 973 National Basic Research Program of China [2012CB114600]
- Project of Science and Technology New Star in Zhu Jiang Guangzhou City [2012J2200083]
Gloverins are basic, glycine-rich and heat-stable antibacterial proteins (similar to 14- kDa) in lepidopteran insects with activity against Escherichia coli, Gram-positive bacteria, fungi and a virus. Hyalophora gloveri gloverin adopts a random coil structure in aqueous solution but has alpha-helical structure in membrane-like environment, and it may interact with the lipid A moiety of lipopolysaccharide (LPS). Manduca sexta gloverin binds to the O-specific antigen and outer core carbohydrate of LPS. In the silkworm Bombyx mori, there are four gloverins with slightly acidic to neutral isoelectric points. In this study, we investigate structural and binding properties and activities of B. mori gloverins (BmGlvs), as well as correlations between structure, binding property and activity. Recombinant BmGlv1-4 were expressed in bacteria and purified. Circular dichroism (CD) spectra showed that all four BmGlvs mainly adopted random coli structure (>50%) in aqueous solution in. regardless of pH, but contained alpha-helical structure in the presence of 1,1,1,3,3,3-hexafluoro-2-propanol (HFIP), smooth and rough mutants (Ra, Rc and Re) of LPS and lipid A. Plate ELISA assay showed that BmGlvs at pH 5.0 bound to rough mutants of LPS and lipid A but not to smooth LPS. Antibacterial activity assay showed that positively charged BmGlvs (at pH 5.0) were active against E. coli mutant strains containing rough LPS but inactive against E. coli with smooth LPS. Our results suggest that binding to rough LPS is the prerequisite for the activity of BmGlvs against E. coli. (C) 2013 Elsevier Ltd.,All rights reserved.
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