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Characterization and assembly of a GFP-tagged cylindriform silk into hexameric complexes.

Öster, Carl LU ; Svensson Bonde, Johan LU ; Bülow, Leif LU and Dicko, Cedric LU (2014) In Biopolymers 101(4). p.378-390
Abstract
Spider silk has been studied extensively for its attractive mechanical properties and potential applications in medicine and industry. The production of spider silk, however, has been lagging behind for lack of suitable systems. Our approach focuses on solving the production of spider silk by designing, expressing, purifying and characterizing the silk from cylindriform glands. We show that the cylindriform silk protein, in contrast to the commonly used dragline silk protein, is fully folded and stable in solution. With the help of GFP as a fusion tag we enhanced the expression of the silk protein in Escherichia coli and could optimize the downstream processing. Secondary structures analysis by circular dichroism and FTIR shows that the... (More)
Spider silk has been studied extensively for its attractive mechanical properties and potential applications in medicine and industry. The production of spider silk, however, has been lagging behind for lack of suitable systems. Our approach focuses on solving the production of spider silk by designing, expressing, purifying and characterizing the silk from cylindriform glands. We show that the cylindriform silk protein, in contrast to the commonly used dragline silk protein, is fully folded and stable in solution. With the help of GFP as a fusion tag we enhanced the expression of the silk protein in Escherichia coli and could optimize the downstream processing. Secondary structures analysis by circular dichroism and FTIR shows that the GFP-Silk fusion protein is predominantly α-helical, and that pH can trigger a α- to β-transition resulting in aggregation. Structural analysis by small angle x-ray scattering suggests that the GFP-Silk exists in the form of a hexamer in solution. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Biopolymers
volume
101
issue
4
pages
378 - 390
publisher
John Wiley & Sons
external identifiers
  • pmid:23955662
  • wos:000336575300008
  • scopus:84897681543
ISSN
0006-3525
DOI
10.1002/bip.22374
language
English
LU publication?
yes
id
f23562af-ef70-446b-9220-5bc82d809d50 (old id 4005559)
date added to LUP
2013-09-09 10:57:52
date last changed
2017-03-12 03:02:12
@article{f23562af-ef70-446b-9220-5bc82d809d50,
  abstract     = {Spider silk has been studied extensively for its attractive mechanical properties and potential applications in medicine and industry. The production of spider silk, however, has been lagging behind for lack of suitable systems. Our approach focuses on solving the production of spider silk by designing, expressing, purifying and characterizing the silk from cylindriform glands. We show that the cylindriform silk protein, in contrast to the commonly used dragline silk protein, is fully folded and stable in solution. With the help of GFP as a fusion tag we enhanced the expression of the silk protein in Escherichia coli and could optimize the downstream processing. Secondary structures analysis by circular dichroism and FTIR shows that the GFP-Silk fusion protein is predominantly α-helical, and that pH can trigger a α- to β-transition resulting in aggregation. Structural analysis by small angle x-ray scattering suggests that the GFP-Silk exists in the form of a hexamer in solution.},
  author       = {Öster, Carl and Svensson Bonde, Johan and Bülow, Leif and Dicko, Cedric},
  issn         = {0006-3525},
  language     = {eng},
  number       = {4},
  pages        = {378--390},
  publisher    = {John Wiley & Sons},
  series       = {Biopolymers},
  title        = {Characterization and assembly of a GFP-tagged cylindriform silk into hexameric complexes.},
  url          = {http://dx.doi.org/10.1002/bip.22374},
  volume       = {101},
  year         = {2014},
}