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Binding of Cu(II)-poly(N-isopropylacrylamide/vinylimidazole) copolymer to histidine-tagged protein: a surface plasmon resonance study.

Kumar, Ashok LU ; Kamihira, M; Galaev, Igor LU ; Iijima, S and Mattiasson, Bo LU (2003) In Langmuir 19(3). p.865-871
Abstract
The thermoresponsive copolymer of N-isopropylacrylamide (NIPAM) with 1-vinylimidazole (VI), poly(NIPAM-VI), synthesized by radical polymerization has been used to purify the histidine-tagged green flourescent protein (His-tag GFP) from recombinant E. coli by metal-chelate affinity precipitation. The purified protein was immobilized on the BIAcore sensor chip by carbodiimide coupling. Affinity binding of the Cu(II)-loaded copolymer, poly(NIPAM-VI), to the His-tag GFP-immobilized surface was monitored by surface-plasmon-resonance (SPR) measurements. Complete recovery of the metal copolymer from the surface was achieved with either using the monomer displacer, 200 mM imidazole buffer, or the polymeric displacer, copolymer of poly(NIPAM-VI)... (More)
The thermoresponsive copolymer of N-isopropylacrylamide (NIPAM) with 1-vinylimidazole (VI), poly(NIPAM-VI), synthesized by radical polymerization has been used to purify the histidine-tagged green flourescent protein (His-tag GFP) from recombinant E. coli by metal-chelate affinity precipitation. The purified protein was immobilized on the BIAcore sensor chip by carbodiimide coupling. Affinity binding of the Cu(II)-loaded copolymer, poly(NIPAM-VI), to the His-tag GFP-immobilized surface was monitored by surface-plasmon-resonance (SPR) measurements. Complete recovery of the metal copolymer from the surface was achieved with either using the monomer displacer, 200 mM imidazole buffer, or the polymeric displacer, copolymer of poly(NIPAM-VI) (26 mol % VI). The conformation of the copolymer was a critical factor for the metal interactions and hence displacement of the metal copolymer. With the proposed conformation of protein-like copolymers (Wahlund, P.-O.; Galaev, I. Yu.; Kazakov, S. A.; Lozinsky, V. I.; Mattiasson, B. Macromol. Biosci. 2002, 2, 33.), the SPR study confirmed the prediction of exposed imidazole groups in the poly(NIPAM-VI). The complete elution of the affinity-bound metal copolymer was achieved with protein-like copolymer (imidazole groups exposed to the outer solution), and no recovery was obtained with IMAC nonbound copolymer fraction (imidazole groups unexposed). The SPR measurement showed a sharp phase transition of affinity adsorbed thermoresponsive Cu(II)-poly(NIPAM-VI) copolymer at 32 C, thus proposing a sensitive way to determine lower critical solution temperature. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
in
Langmuir
volume
19
issue
3
pages
865 - 871
publisher
The American Chemical Society
external identifiers
  • wos:000180737100059
  • scopus:0037418008
ISSN
0743-7463
DOI
10.1021/la020669e
language
English
LU publication?
yes
id
9cbcf0d3-16b6-4148-91a7-3aaa3e0007bf (old id 129168)
date added to LUP
2007-07-02 12:41:20
date last changed
2018-10-03 09:30:52
@article{9cbcf0d3-16b6-4148-91a7-3aaa3e0007bf,
  abstract     = {The thermoresponsive copolymer of N-isopropylacrylamide (NIPAM) with 1-vinylimidazole (VI), poly(NIPAM-VI), synthesized by radical polymerization has been used to purify the histidine-tagged green flourescent protein (His-tag GFP) from recombinant E. coli by metal-chelate affinity precipitation. The purified protein was immobilized on the BIAcore sensor chip by carbodiimide coupling. Affinity binding of the Cu(II)-loaded copolymer, poly(NIPAM-VI), to the His-tag GFP-immobilized surface was monitored by surface-plasmon-resonance (SPR) measurements. Complete recovery of the metal copolymer from the surface was achieved with either using the monomer displacer, 200 mM imidazole buffer, or the polymeric displacer, copolymer of poly(NIPAM-VI) (26 mol % VI). The conformation of the copolymer was a critical factor for the metal interactions and hence displacement of the metal copolymer. With the proposed conformation of protein-like copolymers (Wahlund, P.-O.; Galaev, I. Yu.; Kazakov, S. A.; Lozinsky, V. I.; Mattiasson, B. Macromol. Biosci. 2002, 2, 33.), the SPR study confirmed the prediction of exposed imidazole groups in the poly(NIPAM-VI). The complete elution of the affinity-bound metal copolymer was achieved with protein-like copolymer (imidazole groups exposed to the outer solution), and no recovery was obtained with IMAC nonbound copolymer fraction (imidazole groups unexposed). The SPR measurement showed a sharp phase transition of affinity adsorbed thermoresponsive Cu(II)-poly(NIPAM-VI) copolymer at 32 C, thus proposing a sensitive way to determine lower critical solution temperature.},
  author       = {Kumar, Ashok and Kamihira, M and Galaev, Igor and Iijima, S and Mattiasson, Bo},
  issn         = {0743-7463},
  language     = {eng},
  number       = {3},
  pages        = {865--871},
  publisher    = {The American Chemical Society},
  series       = {Langmuir},
  title        = {Binding of Cu(II)-poly(N-isopropylacrylamide/vinylimidazole) copolymer to histidine-tagged protein: a surface plasmon resonance study.},
  url          = {http://dx.doi.org/10.1021/la020669e},
  volume       = {19},
  year         = {2003},
}