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Recombinant expression and characterization of an epididymis-specific antimicrobial peptide BIN1b/SPAG11E

Paper ID Volume ID Publish Year Pages File Format Full-Text
24753 43535 2009 5 PDF Available
Title
Recombinant expression and characterization of an epididymis-specific antimicrobial peptide BIN1b/SPAG11E
Abstract

BIN1b was reported as an 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 β-sheet. The 2D 15N-1H heteronuclear single-quantum coherence (HSQC) spectra suggested that the fusion protein adopted a compact three-dimensional structure at pH 4.5. Colony forming unit (CFU) inhibition assay demonstrated that 25 μM fusion protein at pH 7.4 had an antimicrobial activity of 40% against E. coli K12D31, 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 on the antimicrobial activity.

Keywords
BIN1b; Size-exclusion; CD; HSQC; Oligomerization; Antimicrobial activity
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Recombinant expression and characterization of an epididymis-specific antimicrobial peptide BIN1b/SPAG11E
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 139, Issue 1, 1 January 2009, Pages 33–37
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us