Dynamic re‐distribution of protein kinase D (PKD) as revealed by a GFP‐PKD fusion protein: dissociation from PKD activation

FEBS Letters

Cantrell, Doreen²  Iglesias, Teresa³  Rozengurt, Enrique¹  Matthews, Sharon² 

[1] Department of Medicine, School of Medicine and Molecular Biology Institute, University of California, 900 Veteran Avenue, Warren Hall, Room 11-124, Los Angeles, CA 90095, USA;Lymphocyte Activation Laboratory, Imperial Cancer Research Fund, Lincoln's Inn Fields, London WC2A 3PX, UK;Molecular Neuropathobiology Laboratory, Imperial Cancer Research Fund, Lincoln's Inn Fields, London WC2A 3PX, UK

关键词: Cysteine-rich domain;    Membrane localization;    Protein kinase C;    Phorbol ester;    CRD;    cysteine-rich domain;    DAG;    diacylglycerol;    DMEM;    Dulbecco's modified Eagle's medium;    GFP;    green fluorescent protein;    PAGE;    polyacrylamide gel electrophoresis;    PBS;    phosphate-buffered saline;    PDB;    phorbol 12;    13-dibutyrate;    PKC;    protein kinase C;    PKD;    protein kinase D;   

DOI  :  10.1016/S0014-5793(99)01090-X

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

Protein kinase D (PKD)/protein kinase Cμ (PKCμ), a serine/threonine protein kinase with distinct structural and enzymological properties, is rapidly activated in intact cells via PKC. The amino-terminal region of PKD contains a cysteine-rich domain (CRD) that directly binds phorbol esters with a high affinity. Here, we show that treatment of transfected RBL 2H3 cells with phorbol 12,13-dibutyrate (PDB) induces a striking CRD-dependent translocation of PKD from the cytosol to the plasma membrane, as shown by real time visualization of a functional green fluorescent protein (GFP)-PKD fusion protein. A single amino acid substitution in the second cysteine-rich motif of PKD (P287G) prevented PDB-induced membrane translocation but did not affect PKD activation. Our results indicate that PKD translocation and activation are distinct processes that operate in parallel to regulate the activity and localization of this enzyme in intact cells.

【 授权许可】

Unknown