Please use this identifier to cite or link to this item: https://doi.org/10.1016/j.bbrc.2007.07.164
DC FieldValue
dc.titleImproving solubility of NR2B amino-terminal domain of N-methyl-d-aspartate receptor expressed in Escherichia coli
dc.contributor.authorNg, F.-M.
dc.contributor.authorSoh, W.
dc.contributor.authorLow, C.-M.
dc.contributor.authorGeballe, M.T.
dc.date.accessioned2011-09-29T05:54:47Z
dc.date.available2011-09-29T05:54:47Z
dc.date.issued2007
dc.identifier.citationNg, F.-M., Soh, W., Low, C.-M., Geballe, M.T. (2007). Improving solubility of NR2B amino-terminal domain of N-methyl-d-aspartate receptor expressed in Escherichia coli. Biochemical and Biophysical Research Communications 362 (1) : 69-74. ScholarBank@NUS Repository. https://doi.org/10.1016/j.bbrc.2007.07.164
dc.identifier.issn0006291X
dc.identifier.issn10902104
dc.identifier.urihttp://scholarbank.nus.edu.sg/handle/10635/27294
dc.description.abstractThe amino-terminal domains (ATDs) of N-methyl-d-aspartate (NMDA) receptors contain binding sites for modulators and may serve as potential drug targets in neurological diseases. Here, three fusion tags (6×His-, GST-, and MBP-) were fused to the ATD of NMDA receptor NR2B subunit (ATD2B) and expressed in Escherichia coli. Each tag's ability to confer enhanced solubility to ATD2B was assessed. Soluble ATD2B was successfully obtained as a MBP fusion protein. Dynamic light scattering revealed the protein (1 mg/ml) exists as monodispersed species at 25 °C. Functional studies using circular dichroism showed that the soluble MBP-ATD2B bound ifenprodil in a dose-dependent manner. The dissociation constants obtained for ifenprodil were similar in the absence (64 nM) and presence (116 nM) of saturating concentration of maltose. Moreover, the yield of soluble MBP-ATD2B is 18 times higher than the refolded 6×His-ATD2B. We have reported a systematic comparison of three different affinity tagging strategies and identified a rapid and efficient method to obtain large amount of ATD2B recombinant protein for biochemical and structural studies. © 2007 Elsevier Inc. All rights reserved.
dc.description.urihttp://libproxy1.nus.edu.sg/login?url=http://dx.doi.org/10.1016/j.bbrc.2007.07.164
dc.sourceScopus
dc.subjectCircular dichroism
dc.subjectEscherichia coli
dc.subjectGlutathione-S-transferase
dc.subjectMaltose binding protein
dc.subjectNMDA receptor
dc.subjectNR2B
dc.typeArticle
dc.contributor.departmentPHARMACOLOGY
dc.description.doi10.1016/j.bbrc.2007.07.164
dc.description.sourcetitleBiochemical and Biophysical Research Communications
dc.description.volume362
dc.description.issue1
dc.description.page69-74
dc.identifier.isiut000249357700012
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