Conformational analyses of a partially-folded bioactive prodomain of human furin

DOIResolve DOI: http://doi.org/10.1002/bip.20748
AuthorSearch for: ; Search for: ; Search for: ; Search for: ; Search for:
TypeArticle
Volume86
Issue4
Pages329344; # of pages: 16
SubjectAmino Acid Motifs; Biotechnology; chemistry; Furin; Hand; Human; Humans; metabolism; Models,Biological; Nuclear Magnetic Resonance,Biomolecular; Peptides; pha; pharmacology; Protease; Protein; Protein Folding; Protein Structure,Secondary; Protein Structure,Tertiary; Solutions; Trifluoroethanol
AbstractThe 81-residue multifunctional prodomain of human furin adopts only a partially-folded conformational state under near physiological conditions. By use of NMR spectroscopy, we demonstrate that the N-terminal residues 1-46 of the prodomain in 50% trifluoroethanol (TFE) populates backbone conformations containing a short helix, a beta-strand and a helix-loop-helix super-secondary structure with elements of tertiary interactions. (15)N NMR relaxation measurements indicate that the helix-loop-helix region has similar motional characteristics in the fast picosecond to nanosecond timescales. On the other hand, the intervening segment (residues 47-65) is predominantly unstructured with a long and highly flexible region surrounding the protease 'activation loop' followed by a partially helical segment in the C-terminal end. Interestingly, the helix-loop-helix 'fold' was found to be populated even when excised out of the full-length prodomain, since a peptide fragment derived from residues Pro16-Arg49 can also form the helix-loop-helix structure in aqueous solution in the absence of TFE. Structure analyses reveal that two helices orient in an antiparallel fashion directed by the sharing of hydrophobic residues involved in helix-capping interactions. Very importantly, a positively-charged Lys residue replacing His43 in the 16-49 fragment imparts stability to the super-secondary structure at both acidic and neutral pH, while a hydrophobic residue Leu at position 43 appears to destabilize the helical conformation in the 31-44 region. As such, this study provides valuable insights into the structural properties of the furin prodomain in relation to its role in the folding of the furin zymogen and its inhibitory action toward furin
Publication date
AffiliationNRC Biotechnology Research Institute; National Research Council Canada
Peer reviewedNo
NRC number47558
NPARC number3538716
Export citationExport as RIS
Report a correctionReport a correction
Record identifierd686677f-bf34-44b1-8aab-b0f7f0068192
Record created2009-03-01
Record modified2016-05-09
Bookmark and share
  • Share this page with Facebook (Opens in a new window)
  • Share this page with Twitter (Opens in a new window)
  • Share this page with Google+ (Opens in a new window)
  • Share this page with Delicious (Opens in a new window)