Rapid evaluation of nickel binding properties of His-tagged lactate dehydrogenases using surface plasmon resonance
(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:
https://lup.lub.lu.se/record/152651
- author
- Bernaudat, Florent LU and Bülow, Leif LU
- organization
- publishing date
- 2005
- 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
- 2016-04-01 16:47:00
- date last changed
- 2022-04-23 00:28:12
@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 < 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.}}, author = {{Bernaudat, Florent and Bülow, Leif}}, issn = {{0021-9673}}, keywords = {{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}}, doi = {{10.1016/j.chroma.2005.01.079}}, volume = {{1066}}, year = {{2005}}, }