Synthesis of the chelator lipid nitrilotriacetic acid ditetradecylamine (NTA-DTDA) and its use with the IAsys biosensor to study receptor-ligand interactions on model membranes

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dc.contributor.authorAltin, Joseph
dc.contributor.authorWhite, Felix
dc.contributor.authorEaston, Christopher
dc.date.accessioned2015-12-08T22:43:19Z
dc.date.issued2001
dc.date.updated2015-12-08T10:39:34Z
dc.description.abstractThis work describes the synthesis and use of the chelator lipid, nitrilotriacetic acid ditetradecylamine (NTA-DTDA). This lipid is readily dispersed in aqueous media, both alone and when mixed with carrier lipids like dimyristoylphosphatidylcholine (DMPC). Fluorescence microscopic examination of membranes deposited from NTA-DTDA-containing liposomes shows that NTA-DTDA mixes uniformly with the carrier lipid, and does not phase separate. NTA-DTDA-membranes deposited onto the sensing surface of IAsys biosensor cuvettes show good stability, permitting use of the biosensor to study protein interactions. Hexahistidine-tagged proteins including recombinant forms of the extracellular regions of murine B7.1 (B7.1-6H) and of the human erythropoietin receptor (EPOR-6H) bind to NTA-DTDA-membranes; the stability of binding is dependent on both protein concentration, and density of NTA-DTDA. Kinetic measurements show that high stability of anchored proteins (t1/2 ∼ 10-20 h, apparent Kd ∼ 1 nM) can be achieved using membranes containing 25 mol% NTA-DTDA, but low levels of bound protein (< 200 arc seconds). The system is used to study the interaction of human EPO with the EPOR anchored onto NTA-DTDA-membranes. In addition to the biological applications reported recently, the results show that NTA-DTDA can be a useful reagent in the study of receptor-ligand interactions.
dc.identifier.issn0005-2736
dc.identifier.urihttp://hdl.handle.net/1885/37225
dc.publisherElsevier
dc.sourceBiochimica et Biophysica Acta: Biomembranes
dc.subjectKeywords: acetic acid derivative; dimyristoylphosphatidylcholine; erythropoietin receptor; hexahistidine; histidine derivative; lipid; liposome; nitrilotriacetic acid ditetradecylamine; unclassified drug; article; artificial membrane; binding affinity; biosensor; c Chelator lipid; Erythropoietin; IAsys biosensor; Membrane; Nitrilotriacetic acid; Receptor-ligand interaction
dc.titleSynthesis of the chelator lipid nitrilotriacetic acid ditetradecylamine (NTA-DTDA) and its use with the IAsys biosensor to study receptor-ligand interactions on model membranes
dc.typeJournal article
local.bibliographicCitation.issue2
local.bibliographicCitation.lastpage148
local.bibliographicCitation.startpage131
local.contributor.affiliationAltin, Joseph, College of Medicine, Biology and Environment, ANU
local.contributor.affiliationWhite, Felix, College of Medicine, Biology and Environment, ANU
local.contributor.affiliationEaston, Christopher, College of Physical and Mathematical Sciences, ANU
local.contributor.authoremailu4235798@anu.edu.au
local.contributor.authoruidAltin, Joseph, u4235798
local.contributor.authoruidWhite, Felix, u9517259
local.contributor.authoruidEaston, Christopher, u9500570
local.description.embargo2037-12-31
local.description.notesImported from ARIES
local.identifier.absfor030503 - Organic Chemical Synthesis
local.identifier.absfor100299 - Environmental Biotechnology not elsewhere classified
local.identifier.ariespublicationu4325460xPUB146
local.identifier.citationvolume1513
local.identifier.doi10.1016/S0005-2736(01)00344-3
local.identifier.scopusID2-s2.0-0035817263
local.identifier.uidSubmittedByu4325460
local.type.statusPublished Version

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