Recruitment of protein kinase D to the trans-Golgi network via the first cysteine-rich domain.
Protein kinase D (PKD) is a cytosolic protein, which upon binding to the trans-Golgi network (TGN) regulates the fission of transport carriers specifically destined to the cell surface. We have found that the first cysteine-rich domain (C1a), but not the second cysteine-rich domain (C1b), is sufficient for the binding of PKD to the TGN. Proline 155 in C1a is necessary for the recruitment of intact PKD to the TGN. Whereas C1a is sufficient to target a reporter protein to the TGN, mutation of serines 744/748 to alanines in the activation loop of intact PKD inhibits its localization to the TGN. Moreover, anti-phospho-PKD antibody, which recognizes only the activated form of PKD, recognizes the TGN-bound PKD. Thus, activation of intact PKD is important for binding to the TGN.
Maeda
Y
Y
Cell and Developmental Biology Department, Division of Biology, University of California San Diego, La Jolla, CA 92093-0347, USA.
Beznoussenko
G V
GV
Van Lint
J
J
Mironov
A A
AA
Malhotra
V
V
D019943:Amino Acid Substitution; D000906:Antibodies / Q000494:pharmacology; D002460:Cell Line; D003545:Cysteine / Q000378:metabolism*; D004789:Enzyme Activation / Q000502:physiology; D017930:Genes, Reporter; D005982:Glutathione Transferase / Q000235:genetics; D006367:HeLa Cells; D006801:Humans; D016297:Mutagenesis, Site-Directed; D010766:Phosphorylation; D011485:Protein Binding / Q000502:physiology; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism*; D017434:Protein Structure, Tertiary / Q000502:physiology; D021381:Protein Transport / Q000502:physiology; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D021601:trans-Golgi Network / Q000378:metabolism*
D000906:Antibodies; D011993:Recombinant Fusion Proteins; D005982:Glutathione Transferase; C089212:protein kinase D; D011493:Protein Kinase C; D003545:Cysteine