Anchoring of histidine-tagged proteins to molecular printboards: self-assembly, thermodynamic modeling, and patterning.
In this paper the multivalent binding of hexahistidine (His6)-tagged proteins to beta-cyclodextrin (beta-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-His6-tagged maltose binding protein (His6-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 beta-CDs through three Ni.4 complexes. This difference is explained by the high effective beta-CD concentration at the surface and is a new example of supramolecular interfacial expression. In a similar adsorption scheme, SPR proved that the alpha-proteasome could be attached to beta-CD SAMs in a specific manner. Patterning through microcontact printing of (His6)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 (His6)4-DsRed-FT is bound strongly through Ni.4 to the molecular printboard.
Ludden
Manon J W
MJ
Molecular Nanofabrication Group, MESA+ Institute for Nanotechnology, University of Twente, PO Box 217, 7500 AE Enschede, The Netherlands.
Mulder
Alart
A
Schulze
Katrin
K
Subramaniam
Vinod
V
Tampé
Robert
R
Huskens
Jurriaan
J
D000327:Adsorption; D001665:Binding Sites; D002352:Carrier Proteins / Q000737:chemistry*; D006639:Histidine / Q000737:chemistry*; D008164:Luminescent Proteins / Q000737:chemistry; D058619:Maltose-Binding Proteins; D008956:Models, Chemical; D008968:Molecular Conformation; D009532:Nickel / Q000737:chemistry; D009571:Nitrilotriacetic Acid / Q000737:chemistry; D009942:Organometallic Compounds / Q000737:chemistry; D010316:Particle Size; D046988:Proteasome Endopeptidase Complex / Q000737:chemistry; D020349:Surface Plasmon Resonance; D013499:Surface Properties; D013816:Thermodynamics; D013997:Time Factors; D047392:beta-Cyclodextrins / Q000737:chemistry
D002352:Carrier Proteins; D008164:Luminescent Proteins; D058619:Maltose-Binding Proteins; D009942:Organometallic Compounds; D047392:beta-Cyclodextrins; C413663:fluorescent protein 583; D006639:Histidine; D009532:Nickel; D046988:Proteasome Endopeptidase Complex; C031215:betadex; D009571:Nitrilotriacetic Acid