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NMR Studies on Nonreceptor Protein Tyrosine Kinase-6 (PTK6). STRUCTRAL BIOLOGY & MOLECULAR BIOPHISICS LAB YONSEI UNIVERSITY CHUNHUA CHI. Representative of PTK6 IHC in breast carcinomas. No staining (normal cell). Light staining (+1). Moderate staining (+2). strong staining (+3).

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NMR Studies on Nonreceptor Protein Tyrosine Kinase-6 (PTK6)


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    1. NMR Studies on Nonreceptor Protein Tyrosine Kinase-6 (PTK6) STRUCTRAL BIOLOGY & MOLECULAR BIOPHISICS LAB YONSEI UNIVERSITY CHUNHUA CHI

    2. Representative of PTK6 IHC in breast carcinomas No staining (normal cell) Light staining (+1) Moderate staining (+2) strong staining (+3) M Aubele,G Auer, and JMS Bartlett etc. British Journal of Cancer (2007) 96, 801 – 807.

    3. PTK6 Protein tyrosine kinases (PTKs) play an important role as regulators of various cellular functions including proliferation, differentiation, and apoptosis. Also are often involved in tumorigenesis through hyper-activation processes . Human PTK6 is also known as Breast cancer Kinase.

    4. Sequence comparison of nonreceptor tyrosine kinase proteins

    5. Cloning of PTK6 445 451 72 78 170 192 11 1 N C SH3 SH2 Kinase Linker SH3 Linker SH3 SH2 Linker

    6. Purification of SH3 domain & backbone assignment

    7. Solution structure of SH3 domain

    8. Solution structure of Linker bounded to SH3 domain

    9. Solution structure of SH3 domain bounded to Linker SH3: LInker Black :1:0 Green:1:1 Blue :1:3 Red :1:5

    10. Structure model of SH3/Linker complex

    11. In vitro kinase assay

    12. Purification of PTK6-SH32L M sup F/T W Elu Cut 21 22 23 kDa 55.4 Absorbance 36.5 31 PTK6-SH32L 21.5 14.4 6 N-NTA Column Work Enzyme treat Gel-filtration Elution Volume

    13. NMR spectra of PTK6-SH32L A B 1H (ppm) 1D proton spectra 1H-15N 2D HSQC

    14. Conclusions Linker region is specific to PTK6 Hydrophobic interactions between SH3 domain and proline residues in the Linker is important for catalytic activity of PTK6 the hydrophobic patch of SH3 domain plays an important role in auto-regulation of PTK6

    15. Acknowledgment Yonsei univ. KIST Prof. Weontae Lee Dr. Sunggeon Ko Dr. Heechul Ann Kyo-eun Ahn Youngnmin Lee