Recombinant expression and characterization of an epididymis-specific antimicrobial peptide BIN1b/SPAG11E
| Autor: | Yandong Lian, Hongchang Gao, Donghai Lin, Chenyun Guo, Heguo Yu, Yonglian Zhang, Hua Diao |
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| Rok vydání: | 2009 |
| Předmět: |
Male
Circular dichroism beta-Defensins Recombinant Fusion Proteins Molecular Sequence Data Size-exclusion chromatography Colony Count Microbial Bioengineering Peptide medicine.disease_cause Applied Microbiology and Biotechnology Protein Structure Secondary law.invention law Escherichia coli medicine Humans Amino Acid Sequence Nuclear Magnetic Resonance Biomolecular Chromatography High Pressure Liquid Epididymis chemistry.chemical_classification Chemistry Circular Dichroism General Medicine Fusion protein Beta defensin Biochemistry Chromatography Gel Recombinant DNA Heteronuclear single quantum coherence spectroscopy Antimicrobial Cationic Peptides Biotechnology |
| Zdroj: | Journal of Biotechnology. 139:33-37 |
| ISSN: | 0168-1656 |
| Popis: | BIN1b was reported as all epididymis-specific beta-defensin antimicrobial peptide. In this paper. the recombinant BIN1b was expressed and purified by fusing with GB1-His tag. The size-exclusion gel filtration experiment indicated that the fusion protein GB1-BIN1b formed multimers at pH 7.4. and existed as monomer at pH 4.5. The oligomerization of GB1-BIN1b was only related to PH value, neither to NaCl concentration nor protein concentration. Far-UV circular dichroism (CD) spectra also showed the fusion protein had more ordered secondary structures at pH 4.5 than at pH 7.4,as a negative peak appeared around 218 nm indicative of typical beta-sheet. The 2D (15)N-(1)H heteronuclear single-quantum coherence (HSQC) spectra suggested that the fusion protein adopted a compact three-dimensional structure at pH 4.5. Colony following unit (CFU) inhibition assay demonstrated that 25 mu M fusion protein at pH 7.4 had all anti microbial activity of 40% against E. coli K(12)D(31). which might imply the fusion protein functions as multimeric states. In conclusion the GB1 fusion partner helps BIN1b form a stable homogenous conformation to facilitate subsequent structural determination Without a significant effect oil the antimicrobial activity. (c) 2008 Elsevier B.V. All rights reserved. |
| Databáze: | OpenAIRE |
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