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NTA‐mediated protein capturing strategy in screening experiments for small organic molecules by surface plasmon resonance
Author(s) -
Yoshitani Naoei,
Saito Kazuki,
Saikawa Wakana,
Asanuma Miwako,
Yokoyama Shigeyuki,
Hirota Hiroshi
Publication year - 2007
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.200600364
Subject(s) - surface plasmon resonance , histidine , protein subunit , chemistry , peptide , histidine kinase , biophysics , target protein , protein–protein interaction , small molecule , plasma protein binding , ligand (biochemistry) , nanotechnology , combinatorial chemistry , biochemistry , materials science , biology , nanoparticle , enzyme , receptor , gene
Nitrilotriacetate (NTA)‐mediated capture of a histidine‐tagged protein is widely used as an easy and simple method to reversibly immobilize the protein onto a sensor chip for surface plasmon resonance (SPR). However, in spite of its advantages, the NTA‐capturing strategy is rarely employed for ligand screening experiments using SPR, because it was thought to cause substantial errors in binding responses, due to the inevitable protein dissociation during the monitoring period. In this study, as demonstrated in a ligand screening for the histidine‐tagged SH3 domain of the human phosphatidylinositol 3‐kinase p85α subunit, false responses after adhesion of undesirable compounds to a target protein could be minimized with the NTA strategy, while binding responses of a positive control peptide still stayed within a 1%‐deviation against the theoretical binding capacity.