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Isolating the whole complex of target proteins of FK506 using affinity resins from novel solid phases
Authors:Teruki Takahashi  Takaaki Shiyama  Tomoko Mori  Ken Hosoya  Akito Tanaka
Institution:(1) Chemistry Department, Reverse Proteomics Research Institute Co., Ltd., 2-6-7 Kazusa-Kamatari, Chiba 292-0818, Japan;(2) Department of Polymer Science and Engineering, Kyoto Institute of Technology, Matsugasaki, Sakyo-ku, Kyoto 606-8585, Japan
Abstract:The development of novel solid phases enabled us to create affinity resins that could be used to isolate the whole complex of target proteins responsible for the immunosuppressive effects of FK506 from rat brain lysate, whereas the affinity resins from commercially available matrices could not achieve this isolation. The results illustrate the enhanced effectiveness of the affinity resin made from this novel material at identifying the target protein of the bioactive compound compared to resins made from the well-known materials Affigel or Toyopearl. This effectiveness arises because the novel material is hydrophilic enough to reduce nonspecific binding proteins and because it has a higher density of ligands that capture the nonubiquitous target protein. Electronic Supplementary Material Supplementary material is available for this article at
Keywords:Affinity resins  Poly(methacrylate)  FK506  Calcineurin  Calmodulin
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