title: The Construction and Functional Study of Protein Kinase Inhibitor Phage.
abstract: A DNA fragment, which encoded the heat-stable protein kinase inhibitor (PKI) (5-24) of camp-dependent protein kinase (cAPK), was synthesized and cloned into phage display vector fe-tet-DOG1. Thus, PKI(5-24) was displayed on the surface of phage fd, which was termed PKI phage (cAPK inhibitor phage), in a form fused with gene III protein (g3p). It was showed that PKI phage not only repressed cAPK effectively, but also bound with the immobilized recombinant His(6)-tag mouse cAPK-Calpha( His(6)-mCalpha) specifically. The bound PKI phages could be quantitatively eluted under acidic conditions. Model affinity screening demonstrated that PKI phages could be selectively enriched from the mixture of PKI phage and wild-type phages (1:10(8)) using affinity chromatography of immobilized His(6)-mCalpha. These results suggest that selecting protein kinase inhibitor by phage display technique is feasible.
authors.affiliation: Shanghai Institute of Biochemistry, the Chinese Academy of Sciences, Shanghai 200031, China.
authors.forename: Chang-Zheng
authors.initials: CZ
authors.lastname: Chen
authors.forename: Xin-Ying
authors.initials: XY
authors.lastname: Yang
authors.forename: Qi-Chang
authors.initials: QC
authors.lastname: Xia
authors.forename: Bo-Liang
authors.initials: BL
authors.lastname: Li
authors.forename: Ying-Lai
authors.initials: YL
authors.lastname: Wang