Isolating the whole complex of target proteins of FK506 using affinity resins from novel solid phases.
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.
Takahashi
Teruki
T
Chemistry Department, Reverse Proteomics Research Institute Co., Ltd., 2-6-7 Kazusa-Kamatari, Chiba, 292-0818, Japan.
Shiyama
Takaaki
T
Mori
Tomoko
T
Hosoya
Ken
K
Tanaka
Akito
A
D000818:Animals; D001921:Brain / Q000378:metabolism; D019703:Calcineurin / Q000302:isolation & purification* / Q000378:metabolism; D002147:Calmodulin / Q000302:isolation & purification* / Q000378:metabolism; D002846:Chromatography, Affinity / Q000295:instrumentation* / Q000379:methods; D015394:Molecular Structure; D051381:Rats; D012117:Resins, Synthetic / Q000737:chemistry*; D013379:Substrate Specificity; D016559:Tacrolimus / Q000378:metabolism*; D022061:Tacrolimus Binding Protein 1A / Q000302:isolation & purification* / Q000378:metabolism; D022021:Tacrolimus Binding Proteins / Q000302:isolation & purification* / Q000378:metabolism
D002147:Calmodulin; D012117:Resins, Synthetic; D019703:Calcineurin; D022061:Tacrolimus Binding Protein 1A; D022021:Tacrolimus Binding Proteins; C403485:tacrolimus binding protein 4; D016559:Tacrolimus