title: Oriented binding of the His6-tagged carboxyl-tail of the L-type Ca2+ channel alpha1-subunit to a new NTA-functionalized self-assembled monolayer.
abstract: Oriented stable binding of functional proteins on surfaces is of fundamental interest for receptor/ligand studies in atomic force microscopy (AFM) and surface plasmon resonance (SPR) experiments. Here we have chosen the His6-tagged carboxyl-tail (C-tail) of the alpha1c-subunit of the L-type Ca2+ channel and calmodulin (CaM) as its cognitive partner as a model system to develop a new functional surface. Covalently attached self-assembled monolayers on ultraflat gold containing NTA-thiols to which the His6-tagged C-tail was bound and thiols with triethylene-glycol groups as matrix-thiols represented the system of choice. The topography of this surface was characterized using AFM; its ability to bind C-tail proteins oriented and stable was confirmed by SPR measurements and by complementary force spectroscopy experiments with a CaM4-construct covalently attached to the tip. The developed anchoring strategy can now be used to study receptor/ligand interactions in general applying force spectroscopy and SPR on His6-tagged proteins oriented immobilized onto this new NTA-functionalized self-assembled monolayer.
authors.affiliation: Institute for Biophysics, University of Linz, Altenbergerstrasse 69, 4020 Linz, Austria.
authors.forename: Roland
authors.initials: R
authors.lastname: Gamsjaeger
authors.forename: Barbara
authors.initials: B
authors.lastname: Wimmer
authors.forename: Heike
authors.initials: H
authors.lastname: Kahr
authors.forename: Ali
authors.initials: A
authors.lastname: Tinazli
authors.forename: Srdjan
authors.initials: S
authors.lastname: Picuric
authors.forename: Suman
authors.initials: S
authors.lastname: Lata
authors.forename: Robert
authors.initials: R
authors.lastname: Tampé
authors.forename: Yves
authors.initials: Y
authors.lastname: Maulet
authors.forename: Hermann J
authors.initials: HJ
authors.lastname: Gruber
authors.forename: Peter
authors.initials: P
authors.lastname: Hinterdorfer
authors.forename: Christoph
authors.initials: C
authors.lastname: Romanin