Expression system reference p16 p16 folding studies

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Expression system Reference p16 p16 Folding studies NMR p16 GST-p16 E. coli (IBs) E. coli (soluble) Tang, 1999 Byeon, 1998 p18 p18 NMR X-ray: 1.95 Å GST-p18 p18 E. coli (soluble) BL21 (soluble) Yuan, 1999 Venkataramani, 1998 p19 NMR p19 E. coli (IBs) Baumgartner, 1999 p19/cdk6, p16/cdk6 p19/cdk6 X-ray: 2.8 Å X-ray: 3.4 Å X-ray: 1.9 Å cdk6 GST-p19/p16 p19 GST-cdk6 Baculo /insect cells E. coli (soluble) E. coli (soluble) Baculo /insect cells Russo, 1998 Brotherton, 1998 p18/cdk6/cycK X-ray: 2.9 Å GST-cycK GST-p18 cdk6 E. coli (soluble) E. coli (soluble) Baculo /insect cells Jeffrey, 2000 cycA-cdk2 cycA-ATP S-cdk2 X-ray: 2.3 Å X-ray: 2.6 Å cdk2 cycA: Baculo /insect cells E.coli (soluble) Jeffery, 1995 Russo, 1996 cycA-ckk2-p27 X-ray: 2.3 Å p27 E. coli (soluble) Russo, 1996 No strcuture GST-cdk4; cdk4 Baculo /insect cells cdk4 (mimic cdk2) X-ray cdk2, engineered cdk4 pocket Baculo /insect cells Ikuta, 2001
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Pharmaceuticals D. Ambrosius; slide 21 Proteine/RAMC-Presentation-9-01 Summary Proteins show a tremendous diversity with respect to - biological function and cellular location - structure, conformation and stability E. coli is a very attractive expression system with respect to time, yield, costs and production of isotope labeled proteins Application of in vitro protein refolding is a powerful tool to generate native structured proteins and should be considered as alternative The protein kinase family is regulated by multiple mechanism and show conformational diversity of catalytic cores; high degree of flexibility - e.g. IRK(3P) and LCK (Tyr kinases) show structural homology to cAPK and cdks (Ser/Thr kinases)
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Pharmaceuticals D. Ambrosius; slide 22 Proteine/RAMC-Presentation-9-01 Acknowledgement PEX: S. Kanzler, H. Brandstetter (MPI) MDM2: G. Saalfrank, Ch. Breitenlechner (MPI), U. Jacob (MPI) IL-16: B. Essig , P. Mühlhahn (MPI), T. Holak (MPI) MIA: G. Saalfrank, C. Hergersberg, R. Stoll (MPI), T. Holak (MPI) cAPK: G. Achhammer, E. Liebig, Ch. Breitenlechner (MPI) cdks: H. Hertenberger, J. Kluge, U. Jucknischke G-CSF: S. Stammler, M. Leidenberger, U. Michaelis, T. Zink (MPI), T. Holak (MPI)
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