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Anchoring of Histidine‐Tagged Proteins to Molecular Printboards: Self‐assembly, Thermodynamic Modeling, and Patterning
Author(s) -
Ludden Ma J. W.,
Mulder Alart,
Schulze Katrin,
Subramaniam Vinod,
Tampé Robert,
Huskens Jurriaan
Publication year - 2008
Publication title -
chemistry – a european journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.687
H-Index - 242
eISSN - 1521-3765
pISSN - 0947-6539
DOI - 10.1002/chem.200701478
Subject(s) - chemistry , ethylene glycol , supramolecular chemistry , crystallography , molecule , self assembly , histidine , linker , monolayer , organic chemistry , biochemistry , computer science , enzyme , operating system
In this paper the multivalent binding of hexahistidine (His 6 )‐tagged proteins to β‐cyclodextrin (β‐CD) self‐assembled monolayers (SAMs) by using the nickel(II) complex of a hetero‐divalent orthogonal adamantyl nitrilotriacetate linker ( 4 ) is described. Nonspecific interactions were suppressed by using monovalent adamantyl‐hexa(ethylene glycol) derivative 3 . With the mono‐His 6 ‐tagged maltose binding protein (His 6 ‐MBP), thermodynamic modeling based on surface plasmon resonance (SPR) titration data showed that the MBP molecules in solution were linked, on average, to Ni ⋅4 in 1:1 stoichiometry. On the surface, however, the majority of His 6 ‐MBP was complexed to surface‐immobilized β‐CDs through three Ni ⋅4 complexes. This difference is explained by the high effective β‐CD concentration at the surface and is a new example of supramolecular interfacial expression. In a similar adsorption scheme, SPR proved that the α‐proteasome could be attached to β‐CD SAMs in a specific manner. Patterning through microcontact printing of (His 6 ) 4 ‐DsRed‐fluorescent timer (DsRed‐FT), which is a tetrameric, visible autofluorescent protein, was carried out in the presence of Ni ⋅4 . Fluorescence measurements showed that the (His 6 ) 4 ‐DsRed‐FT is bound strongly through Ni ⋅4 to the molecular printboard.