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  Molecular recognition of histidine-tagged molecules by metal-chelating lipids monitored by fluorescence energy transfer and correlation spectroscopy

Dorn, I. T., Neumaier, K. R., & Tampe, R. (1998). Molecular recognition of histidine-tagged molecules by metal-chelating lipids monitored by fluorescence energy transfer and correlation spectroscopy. Journal of the American Chemical Society, 120(12), 2753-2763.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0010-721E-B Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0010-721F-9
Genre: Journal Article

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Dorn, I. T., Author
Neumaier, K. R., Author
Tampe, R.1, Author
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1External Organizations, ou_persistent22              

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Free keywords: Surface-plasmon resonance; Air-water-interface; Reversible immobilization; Recombinant proteins; Membranes; Monolayers; Binding; Crystallization; Coordination; Bilayers.; Chemistry. Chemistry & analysis.
 Abstract: Complex binding of proteins by metal-chelating Lipids via surface-exposed or protein-engineered histidines provides an universal and powerful concept for the orientation and two-dimensional crystallization of proteins at self-organized interfaces. To demonstrate pair formation between individual histidine-tagged molecules and chelator lipids on the molecular level, we have synthesized novel lipids bearing both a Ni-NTA chelator and a fluorescent group. These lipids serve as spectroscopic probes to visualize directly the molecular recognition of fluorescence-labeled histidine-tagged peptides by metal-chelating lipids using fluorescence resonance energy transfer (FRET). The molecular docking to chelator lipids assembled in mono- or bilayers is highly specific, revealing only 3% unspecific adsorption and a binding constant of 3 mu M. The affinity constant was confirmed by fluorescence correlation spectroscopy (FCS) on single molecules, where the ratio of lipid-bound to free was analyzed by their intrinsically different diffusion times passing through a confocal volume of about 1 fL. By using a model peptide most of the electrostatic and steric contribution to the binding process can be neglected. Therefore, the affinity constant can serve as a standard value for the binding of histidine-tagged proteins to chelator lipid interfaces. [References: 51]

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 Dates: 1998-04-01
 Publication Status: Published in print
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 Identifiers: eDoc: 318465
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Title: Journal of the American Chemical Society
Source Genre: Journal
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Pages: - Volume / Issue: 120 (12) Sequence Number: - Start / End Page: 2753 - 2763 Identifier: -