Premium
High‐Affinity Chelator Thiols for Switchable and Oriented Immobilization of Histidine‐Tagged Proteins: A Generic Platform for Protein Chip Technologies
Author(s) -
Tinazli Ali,
Tang Jilin,
Valiokas Ramûnas,
Picuric Srdjan,
Lata Suman,
Piehler Jacob,
Liedberg Bo,
Tampé Robert
Publication year - 2005
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.200500154
Subject(s) - chemistry , histidine , surface plasmon resonance , biophysics , chelation , proteome , protein–protein interaction , self assembled monolayer , nanotechnology , monolayer , combinatorial chemistry , materials science , nanoparticle , biochemistry , enzyme , organic chemistry , biology
Abstract Protein micro‐/nanoarrays are becoming increasingly important in systematic approaches for the exploration of protein–protein interactions and dynamic protein networks, so there is a high demand for specific, generic, stable, uniform, and locally addressable protein immobilization on solid supports. Here we present multivalent metal‐chelating thiols that are suitable for stable binding of histidine‐tagged proteins on biocompatible self‐assembled monolayers (SAMs). The architectures and physicochemical properties of these SAMs have been probed by various surface‐sensitive techniques such as contact angle goniometry, ellipsometry, and infrared reflection–absorption spectroscopy. The specific molecular organization of proteins and protein complexes was demonstrated by surface plasmon resonance, confocal laser scanning, and atomic force microscopy. In contrast to the mono‐NTA/His 6 tag interaction, which has major drawbacks because of its low affinity and fast dissociation, drastically improved stability of protein binding by these multivalent chelator surfaces was observed. The immobilized histidine‐tagged proteins are uniformly oriented and retain their function. At the same time, proteins can be removed from the chip surface under mild conditions (switchability). This new platform for switchable and oriented immobilization should assist proteome‐wide wide analyses of protein–protein interactions as well as structural and single‐molecule studies.