Advanced

Rapid evaluation of nickel binding properties of His-tagged lactate dehydrogenases using surface plasmon resonance

Bernaudat, Florent LU and Bülow, Leif LU (2005) In Journal of Chromatography A 1066(1-2). p.219-224
Abstract
The use of surface plasmon resonance (SPR), for the comparison of metal binding properties of polyhistidine tags, was evaluated. Six different tags containing various number of histidines, either none (tags n and t), three (tags H(3)A(3) and H(2)AHA(2)H) or six (tags H-6 and HiS(6)) were genetically fused to the N-terminal of lactate dehydrogenase (LDH). The binding ability of these constructs to nickel ions, immobilised with nitrilotriacetic acid (NTA), was tested both by conventional immobilised metal ion affinity chromatography (IMAC) and SPR. The relative binding strengths of the tags to nickel were identical using both methods (n approximate to t < HA(2)HA(2)H < H(3)A(3) < HiS(6) < H-6) confirming the value of the SPR... (More)
The use of surface plasmon resonance (SPR), for the comparison of metal binding properties of polyhistidine tags, was evaluated. Six different tags containing various number of histidines, either none (tags n and t), three (tags H(3)A(3) and H(2)AHA(2)H) or six (tags H-6 and HiS(6)) were genetically fused to the N-terminal of lactate dehydrogenase (LDH). The binding ability of these constructs to nickel ions, immobilised with nitrilotriacetic acid (NTA), was tested both by conventional immobilised metal ion affinity chromatography (IMAC) and SPR. The relative binding strengths of the tags to nickel were identical using both methods (n approximate to t < HA(2)HA(2)H < H(3)A(3) < HiS(6) < H-6) confirming the value of the SPR technique for investigating metal-protein interactions. Protein modelling has also proved to be useful in supporting the experimental results. (Less)
Please use this url to cite or link to this publication:
author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
nitrilotriacetic acid, surface plasmon resonance, affinity tags, immobilised metal ion affinity chromatography, nickel, lactate dehydrogenase
in
Journal of Chromatography A
volume
1066
issue
1-2
pages
219 - 224
publisher
Elsevier
external identifiers
  • wos:000227503600027
  • pmid:15794574
  • scopus:14144250674
ISSN
0021-9673
DOI
10.1016/j.chroma.2005.01.079
language
English
LU publication?
yes
id
fee23d38-33b5-49bb-806e-3ee719a66b47 (old id 152651)
date added to LUP
2007-07-16 08:24:07
date last changed
2017-02-12 04:12:08
@article{fee23d38-33b5-49bb-806e-3ee719a66b47,
  abstract     = {The use of surface plasmon resonance (SPR), for the comparison of metal binding properties of polyhistidine tags, was evaluated. Six different tags containing various number of histidines, either none (tags n and t), three (tags H(3)A(3) and H(2)AHA(2)H) or six (tags H-6 and HiS(6)) were genetically fused to the N-terminal of lactate dehydrogenase (LDH). The binding ability of these constructs to nickel ions, immobilised with nitrilotriacetic acid (NTA), was tested both by conventional immobilised metal ion affinity chromatography (IMAC) and SPR. The relative binding strengths of the tags to nickel were identical using both methods (n approximate to t &lt; HA(2)HA(2)H &lt; H(3)A(3) &lt; HiS(6) &lt; H-6) confirming the value of the SPR technique for investigating metal-protein interactions. Protein modelling has also proved to be useful in supporting the experimental results.},
  author       = {Bernaudat, Florent and Bülow, Leif},
  issn         = {0021-9673},
  keyword      = {nitrilotriacetic acid,surface plasmon resonance,affinity tags,immobilised metal ion affinity chromatography,nickel,lactate dehydrogenase},
  language     = {eng},
  number       = {1-2},
  pages        = {219--224},
  publisher    = {Elsevier},
  series       = {Journal of Chromatography A},
  title        = {Rapid evaluation of nickel binding properties of His-tagged lactate dehydrogenases using surface plasmon resonance},
  url          = {http://dx.doi.org/10.1016/j.chroma.2005.01.079},
  volume       = {1066},
  year         = {2005},
}