Protein Kinase C Phosphorylates Protein Kinase D Activation Loop Ser744 and Ser748 and Releases Autoinhibition by the Pleckstrin Homology Domain

Waldron, Richard T.; Rozengurt, Enrique

doi:10.1074/jbc.m208075200

Cited by 186 publications

(204 citation statements)

References 64 publications

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“…GST or GST-c-jun fusion proteins (4 μg/time point) were phosphorylated by GST-PKD (90 ng/time point), in the presence of PS/PDB vesicles [7] and ATP (100 μM, with 2 μCi/time point [γ-32 P]ATP). SDS-PAGE gels were stained with Coomassie blue and dried, and c-jun fusion protein phosphorylation quantitated with a scintillation counter.…”

Section: Gst-c-jun Fusion Protein and C-jun Peptide Phosphorylation Rmentioning

confidence: 99%

“…PKDs, as well as several protein kinase C (PKC) isoforms, are directly stimulated by the second messenger diacylglycerol. Further, PKDs have emerged as effectors of novel PKC isoforms including PKCε and PKCη, which directly phosphorylate and activate PKDs in novel protein kinase cascades [6,7]. PKD, activated via PKC-mediated activation loop phosphorylation, exhibits dynamic spatiotemporal behavior including nucleocytoplasmic shuttling [6].…”

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confidence: 99%

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Identification of a novel phosphorylation site in c-jun directly targeted in vitro by protein kinase D

Waldron

Whitelegge

Faull

2007

Biochemical and Biophysical Research Communications

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Protein kinase D (PKD) phosphorylates the c-jun amino-terminal in vitro at site(s) distinct from JNK [C. Hurd, R.T. Waldron, E. Rozengurt, Protein kinase D complexes with c-jun N-terminal kinase via activation loop phosphorylation and phosphorylates the c-jun N-terminus, Oncogene 21 (2002) 2154-2160], but the sites have not been identified. Here, metabolic 32 P-labeling of c-jun protein in COS-7 cells indicated that PKD phosphorylates c-jun in vivo at a site(s) between aa 43-93, a region containing important functional elements. On this basis, the PKD-mediated phosphorylation site(s) was further characterized in vitro using GST-c-jun fusion proteins. PKD did not incorporate phosphate into Ser63 and Ser73, the JNK sites in GST-c-jun(1-89). Rather, PKD and JNK could sequentially phosphorylate distinct site(s) simultaneously. By mass spectrometry of tryptic phosphopeptides, Ser58 interposed between the JNK-binding portion of the delta domain and the adjacent TAD1 was identified as a prominent site phosphorylated in vitro by PKD. These data were further supported by kinase reactions using truncations or point-mutations of GST-c-jun. Together, these data suggest that PKD-mediated phosphorylation modulates c-jun at the level of its N-terminal functional domains.

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Section: Gst-c-jun Fusion Protein and C-jun Peptide Phosphorylation Rmentioning

confidence: 99%

mentioning

confidence: 99%

Identification of a novel phosphorylation site in c-jun directly targeted in vitro by protein kinase D

Waldron

Whitelegge

Faull

2007

Biochemical and Biophysical Research Communications

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“…For example, G-proteincoupled receptors or receptor tyrosine kinases that activate PLC and PKCε or PKCη cause the phosphorylation of PKD at Ser744 and Ser748 in the activation loop [16][17][18][19] . Second, Gβγ subunits directly activate PKD1 [20][21][22] .…”

Section: The Pkd Family Belongs To the Camk Groupmentioning

confidence: 99%

“…Unlike other PH domains that bind to lipids, the PH domain in PKD1 can bind to several proteins. Mutations in the PH domain can activate PKD1 [17,24,25] . Deletion of the two zinc-fi ngers also fully activates PKD1 [26,27] .…”

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confidence: 99%

Protein kinase D: a new player among the signaling proteins that regulate functions in the nervous system

Wang

2014

Neurosci. Bull.

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Protein kinase D (PKD) is an evolutionarily-conserved family of protein kinases. It has structural, regulatory, and enzymatic properties quite different from the PKC family. Many stimuli induce PKD signaling, including G-protein-coupled receptor agonists and growth factors. PKD1 is the most studied member of the family. It functions during cell proliferation, differentiation, secretion, cardiac hypertrophy, immune regulation, angiogenesis, and cancer. Previously, we found that PKD1 is also critically involved in pain modulation. Since then, a series of studies performed in our lab and by other groups have shown that PKDs also participate in other processes in the nervous system including neuronal polarity establishment, neuroprotection, and learning. Here, we discuss the connections between PKD structure, enzyme function, and localization, and summarize the recent fi ndings on the roles of PKD-mediated signaling in the nervous system. Keywords: PKD; neuronal polarity; pain modulation; neuroprotection; learning IntroductionMembers of the protein kinase D (PKD) family are diacylglycerol (DAG) and protein kinase C (PKC) effectors.They are activated by the actions of hormones, growth factors, neurotransmitters, and other stimuli through phospholipase C (PLC) [1] . Three widely-expressed mammalian homologs are PKD1 (mouse PKD, human PKCμ) [2,3] , PKD2 [3] , and PKD3 [4] (also named PKC), but the levels of individual PKDs vary in different tissues.Recent fi ndings have revealed that PKDs participate in the regulation of Golgi function through modulating the fi ssion of vesicles from the trans-Golgi network (TGN) [5,6] . Other recent reports have shown that PKDs function during cell proliferation and apoptosis, carcinogenesis, and intracellular trafficking. Here, we describe the connections between PKD structure, enzymatic function, and localization, and then summarize recent fi ndings on the roles of PKD in the nervous system. The PKD Family Belongs to the CaMK GroupThe PKD family comprises PKD1, PKD2, and PKD3. PKD was initially described as an atypical isoform of the PKC family [7] , which is a member of the protein kinase A, G, and C (AGC) serine/threonine kinase subfamily [8,9] . However, later studies revealed that PKD has mixed features of different subclasses of the PKC family, so it does not belong to any one of them. For example, its pleckstrin-homology (PH) domain is closely related to the PKB and G-proteincoupled receptor kinase (GRK) families and is not found in any PKC enzyme, while the cysteine-rich domains are more reminiscent of classical and novel PKCs. The structure and function of the catalytic domain of PKD are quite different from those of the AGC/PKC family members [2][3][4]10] . Indeed, PKD has now been classified as a new family within the (Fig. 1). The elaborate constitution of PKD1 is intricately linked to its catalytic functions, regulation, and intracellular localization (Table 1). PKD1 can be activated through different pathways.First, some stimuli activate PLC, which induces PKD phosp...

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“…The N‐terminal regulatory region encompasses an alanine/proline rich region (AP region), a tandem C1 domain binding diacylglycerol (DAG) and phorbol esters, an oligomerization domain, and a pleckstrin homology (PH) domain 24, 25, 26, 27, 28. Deletion mutagenesis approaches have indicated that the C1 and PH domains have an autoinhibitory effect on kinase activity, which is alleviated by activation loop Ser‐738/742 phosphorylation (hPKD1 numbering) 24, 25, 26. These phosphorylations are exerted by several PKCs, but PKDs can also autophosphorylate on Ser‐738 and Ser‐742 in vitro , and on Ser‐742 in cells 29, 30.…”

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confidence: 99%

Protein kinase D displays intrinsic Tyr autophosphorylation activity: insights into mechanism and regulation

et al. 2018

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The protein kinase D (PKD) family is regulated through multi‐site phosphorylation, including autophosphorylation. For example, PKD displays in vivo autophosphorylation on Ser‐742 (and Ser‐738 in vitro) in the activation loop and Ser‐910 in the C‐tail (hPKD1 numbering). In this paper, we describe the surprising observation that PKD also displays in vitro autocatalytic activity towards a Tyr residue in the P + 1 loop of the activation segment. We define the molecular determinants for this unusual activity and identify a Cys residue (C705 in PKD1) in the catalytic loop as of utmost importance. In cells, PKD Tyr autophosphorylation is suppressed through the association of an inhibitory factor. Our findings provide important novel insights into PKD (auto)regulation.

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Protein Kinase C Phosphorylates Protein Kinase D Activation Loop Ser744 and Ser748 and Releases Autoinhibition by the Pleckstrin Homology Domain

Cited by 186 publications

References 64 publications

Identification of a novel phosphorylation site in c-jun directly targeted in vitro by protein kinase D

Identification of a novel phosphorylation site in c-jun directly targeted in vitro by protein kinase D

Protein kinase D: a new player among the signaling proteins that regulate functions in the nervous system

Protein kinase D displays intrinsic Tyr autophosphorylation activity: insights into mechanism and regulation

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