Phosphatidyl-L-serine Is Necessary for Protein Kinase C's High-Affinity Interaction with Diacylglycerol-Containing Membranes

Newton, Alexandra C.; Keranen, Lisa M.

doi:10.1021/bi00187a035

Cited by 132 publications

(143 citation statements)

References 39 publications

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“…Similar unspecific ionic interaction of the PH domain of ARNO (46) with membrane PS has been reported. Also, the involvement of PS in other signaling pathways such as Raf-1 and PKC activation (21,47,48) has been documented, and the importance of the PS as the potential switch for the GTPase substrate preference for GTPase activating protein has been recently demonstrated (49). The interaction between PS and Akt was supported by the abolition or significant reduction of translocation after the mutation of the basic residues, which have been suggested to interact with negatively charged membrane surfaces (39).…”

Section: Discussionmentioning

confidence: 97%

Docosahexaenoic acid: A positive modulator of Akt signaling in neuronal survival

Akbar

Calderón

Zhao

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

408

319

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Phosphatidylinositol 3-kinase [PI (3)K]/Akt signaling is a critical pathway in cell survival. Here, we demonstrate a mechanism where membrane alteration by the n-3 fatty acid status affects Akt signaling, impacting neuronal survival. Docosahexaenoic acid (DHA), an n-3 polyunsaturated fatty acid highly enriched in neuronal membranes, promotes neuronal survival by facilitating membrane translocation/activation of Akt through its capacity to increase phosphatidylserine (PS), the major acidic phospholipid in cell membranes. The activation of PI (3)K and phosphatidylsinositol triphosphate formation were not affected by DHA, indicating that membrane interaction of Akt is the event responsible for the DHA effect. Docosapentaenoic acid, which replaces DHA during n-3 fatty acid deficiency, was less effective in accumulating PS and translocating Akt and thus less effective in preventing apoptosis. Consistently, in vivo reduction of DHA by dietary depletion of n-3 fatty acids decreased hippocampal PS and increased neuronal susceptibility to apoptosis in cultures. This mechanism may contribute to neurological deficits associated with n-3 fatty acid deficiency and support protective effects of DHA in pathological models such as brain ischemia or Alzheimer's disease

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Section: Discussionmentioning

confidence: 97%

Docosahexaenoic acid: A positive modulator of Akt signaling in neuronal survival

Akbar

Calderón

Zhao

et al. 2005

Proc. Natl. Acad. Sci. U.S.A.

408

319

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“…Values of EC 50 % for reconstitution of binding to PKC␦ were 10.9 Ϯ 0.9% (n ϭ 3) in the absence of calcium and 9.6 Ϯ 1.1% (n ϭ 3) in the presence of calcium. As suggested by Newton and Keranen (37), calcium increases the affinity of PKC for the acidic phospholipids. Our results support that concept but also suggest that the calcium effect is limited to calcium-dependent PKCs rather than to other calcium-independent phorbol ester receptors, such as the novel PKCs and chimaerins.…”

Section: ␤2-chimaerin As a Phorbol Ester Receptormentioning

confidence: 72%

β2-Chimaerin Is a High Affinity Receptor for the Phorbol Ester Tumor Promoters

Caloca

Fernández

Lewin

et al. 1997

Journal of Biological Chemistry

113

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␤2-chimaerin, a member of the GTPase-activating proteins for the small GTP-binding protein p21Rac, possesses a single cysteine-rich domain with high homology to those implicated in phorbol ester and diacylglycerol binding in protein kinase C (PKC) isozymes. We have expressed ␤2-chimaerin in Sf9 insect cells using the baculovirus expression system and determined that, like PKCs, ␤2-chimaerin binds phorbol esters with high affinity in the presence of phosphatidylserine as a cofactor. Scatchard plot analysis using the radioligand [ 3 H]phorbol 12,13-dibutyrate revealed a dissociation constant of 1.9 ؎ 0.2 nM for ␤2-chimaerin. Likewise, ␤2-chimaerin is a high affinity receptor for the bryostatins, a class of atypical PKC activators. A detailed comparison of structure-activity relations using several phorbol ester analogs revealed striking differences in binding recognition between ␤2-chimaerin and PKC␣. Although the diacylglycerol 1-oleoyl-2-acetylglycerol binds with similar potency to both ␤2-chimaerin and PKC␣, the mezerein analog thymeleatoxin has 56-fold less affinity for binding to ␤2-chimaerin. To establish whether ␤2-chimaerin responds to phorbol esters in cellular systems, we overexpressed ␤2-chimaerin in COS-7 cells and monitored its subcellular distribution after phorbol ester treatment. Interestingly, as described previously for PKC isozymes, ␤2-chimaerin translocates from cytosolic to particulate fractions as a consequence of phorbol ester treatment. Our results demonstrate that ␤2-chimaerin is a novel target for the phorbol ester tumor promoters. The expansion of the family of phorbol ester receptors strongly suggests a potential for the "nonkinase" receptors as cellular mediators of the phorbol ester responses.The phorbol esters and related diterpenes are natural compounds that are used widely as tumor promoters in animal models. The search for receptors for the phorbol esters led to the identification of protein kinase C (PKC) 1 as their target (1-5). The complexity and heterogeneity in the biology of phorbol esters suggested the existence of multiple receptors, and 11 PKC isoforms have been identified so far: classic or calciumdependent isozymes (PKC ␣, ␤1, ␤2, and ␥), novel or calciumindependent isozymes (PKC ␦, ⑀, , , and ), and atypical isozymes (PKC and ) (6, 7). The last group is unresponsive to the phorbol esters. DAG, the postulated endogenous activator of PKC, competes with phorbol esters for binding to PKC in in vitro assays, although its binding potency is lower than that of the most commonly used phorbol esters (8). The cysteine-rich domains present at the NH 2 -terminal region of the PKCs are the binding sites for DAG/phorbol esters (9 -13). Each of these 50-or 51-amino acid domains possesses the motif HX 12 CX 2 CX 13/14 CX 2 CX 4 HX 2 CX 7 C, where H is histidine, C is cysteine, and X is any other amino acid. This motif is duplicated in tandem in both the classic and novel PKCs and is present only once in the atypical PKCs. The cysteine-rich domains have been implicated in the associati...

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“…Binding and Activation of PKC␦-It has been long known that among different anionic phospholipids PS specifically enhances the membrane affinity and activity of PKCs (50,51). Recently, however, it has been recognized that PS specificity can vary significantly among PKC isoforms depending on their structures and activation mechanisms.…”

Section: Phosphatidylserine (Ps)-specific Membranementioning

confidence: 99%

Mechanism of Diacylglycerol-induced Membrane Targeting and Activation of Protein Kinase Cδ

Stahelin

Digman

Medkova³

et al. 2004

Journal of Biological Chemistry

121

155

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The regulatory domains of novel protein kinases C (PKC) contain two C1 domains (C1A and C1B), which have been identified as the interaction site for sn-1,2-diacylglycerol (DAG) and phorbol ester, and a C2 domain that may be involved in interaction with lipids and/or proteins. Although recent reports have indicated that C1A and C1B domains of conventional PKCs play different roles in their DAG-mediated membrane binding and activation, the individual roles of C1A and C1B domains in the DAG-mediated activation of novel PKCs have not been fully understood. In this study, we determined the roles of C1A and C1B domains of PKC␦ by means of in vitro lipid binding analyses and cellular protein translocation measurements. Isothermal titration calorimetry and surface plasmon resonance measurements showed that isolated C1A and C1B domains of PKC␦ have opposite affinities for DAG and phorbol ester; i.e. the C1A domain with high affinity for DAG and the C1B domain with high affinity for phorbol ester. Furthermore, in vitro activity and membrane binding analyses of PKC␦ mutants showed that the C1A domain is critical for the DAG-induced membrane binding and activation of PKC␦. The studies also indicated that an anionic residue, Glu 177 , in the C1A domain plays a key role in controlling the DAG accessibility of the conformationally restricted C1A domain in a phosphatidylserine-dependent manner. Cell studies with enhanced green fluorescent protein-tagged PKC␦ and mutants showed that because of its phosphatidylserine specificity PKC␦ preferentially translocated to the plasma membrane under the conditions in which DAG is randomly distributed among intracellular membranes of HEK293 cells. Collectively, these results provide new insight into the differential roles of C1 domains in the DAG-induced membrane activation of PKC␦ and the origin of its specific subcellular localization in response to DAG.

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Phosphatidyl-L-serine Is Necessary for Protein Kinase C's High-Affinity Interaction with Diacylglycerol-Containing Membranes

Cited by 132 publications

References 39 publications

Docosahexaenoic acid: A positive modulator of Akt signaling in neuronal survival

Docosahexaenoic acid: A positive modulator of Akt signaling in neuronal survival

β2-Chimaerin Is a High Affinity Receptor for the Phorbol Ester Tumor Promoters

Mechanism of Diacylglycerol-induced Membrane Targeting and Activation of Protein Kinase Cδ

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