Crystal structure of the C2 domain from protein kinase C-δ

Pappa, Helen S.; Murray‐Rust, Judith; Dekker, Lodewijk V.; Parker, P J; McDonald, N.Q.

doi:10.1016/s0969-2126(98)00090-2

Cited by 111 publications

(86 citation statements)

References 41 publications

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“…It is interesting that the 60% of β-sheet calculated for PKCε correlates also well with the percentages calculated by X-ray diffraction for C2 domains of the same topology, such as PLCδ (53%), cPLA2 (60%) and PKCδ-C2 (53%) domains [17,36,44]. Recent studies have shown the crystal structure of the PKCδ-C2 domain, which belongs to the group of novel PKCs.…”

Section: Structure Of the Pkcε-c2 Domainsupporting

confidence: 61%

“…Recent studies have shown the crystal structure of the PKCδ-C2 domain, which belongs to the group of novel PKCs. However, the low sequence homology between PKCε and PKCδ C2 domains suggests that further structural variations are expected to be found in this (A) (B) group [17]. For example, there is an extended sequence in the CBR1-like loop of PKCε that could adopt a large loop conformation similar to that described in the CBR1 of PLCδ1, PTEN or PI3K [13,34,36].…”

Section: Structure Of the Pkcε-c2 Domainmentioning

confidence: 96%

“…X-ray diffraction analysis of several C2 domains has revealed that the structure consists of a compact β-sandwich composed of two four-stranded β-sheets [1,11,17,36,42,44,47,48]. Basically, three loops at the top of the domain and four at the bottom connect the eight β-strands and, interestingly, two distinct but easily interconverted topological folds have been found: topology I becomes topology II when its N-and C-termini are fused and new termini are generated by cutting the loop between strands β1 and β2 [11,26].…”

Section: Introductionmentioning

confidence: 99%

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Structural Characterization of the C2 Domains of Classical Isozymes of Protein Kinase C and Novel Protein Kinase Cε by using Infrared Spectroscopy

Corbalán-Garcı́a

Garcı́a-Garcı́a

Sánchez-Carrillo

et al. 2003

Journal of Spectroscopy

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Abstract. The amide I regions in the original infrared spectra of PKCα-C2 in the Ca 2+ -free and Ca 2+ -bound states are both consistent with a predominantly β-sheet secondary structure. Spectroscopic studies of the thermal denaturation revealed that for the PKCα-C2 domain alone the secondary structure abruptly changed at 50 • C. While in the presence of Ca 2+ , the thermal stability of the protein increased considerably. Phosphatidic acid binding to the PKCα-C2 domain was characterized, and the lipid-protein binding becoming Ca 2+ -independent when 100 mol% phosphatidic acid vesicles was used. The effect of lipid binding on secondary structure and thermal stability was also studied. In addition, the secondary structure of the C2 domain from the novel PKCε was also determined by IR spectroscopy and β-sheet was seen to be the major structural component. Spectroscopic studies of the thermal denaturation in D2O showed a broadening in the amide I band starting at 45 • C. Phosphatidic acid containing vesicles were used to characterize the effect of lipid binding on the secondary structure. It was observed through thermal stability experiments that the secondary structure did not change upon lipid binding and the protein stability was very high with no significant changes occurring in the secondary structure after heating.

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Section: Structure Of the Pkcε-c2 Domainsupporting

confidence: 61%

Section: Structure Of the Pkcε-c2 Domainmentioning

confidence: 96%

Section: Introductionmentioning

confidence: 99%

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Structural Characterization of the C2 Domains of Classical Isozymes of Protein Kinase C and Novel Protein Kinase Cε by using Infrared Spectroscopy

Corbalán-Garcı́a

Garcı́a-Garcı́a

Sánchez-Carrillo

et al. 2003

Journal of Spectroscopy

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“…The domain consists of an anti-parallel ␤-sandwich composed of two four-stranded sheets. The overall fold of the cPLA 2 C2 domain is similar to the C2 domains from other proteins: synaptotagmin I (SytI-C2A) (21), phospholipase C␦1 (22), protein kinase C␦ (23), and protein kinase C␤ (24). The topology of the domain is identical to that of phospholipase C␦1 and the recently reported calcium-independent C2 domain from protein kinase C␦ (23), whereas it is a circular permutation of the SytI-C2A and protein kinase C␤ topology.…”

mentioning

confidence: 86%

Mapping the Phospholipid-binding Surface and Translocation Determinants of the C2 Domain from Cytosolic Phospholipase A2

Perišić¹,

Paterson²,

Mosedale³

et al. 1999

Journal of Biological Chemistry

120

112

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Cytosolic phospholipase A 2 (cPLA 2 ) plays a key role in the generation of arachidonic acid, a precursor of potent inflammatory mediators. Intact cPLA 2 is known to translocate in a calcium-dependent manner from the cytosol to the nuclear envelope and endoplasmic reticulum. We show here that the C2 domain of cPLA 2 alone is sufficient for this calcium-dependent translocation in living cells. We have identified sets of exposed hydrophobic residues in loops known as calcium-binding region (CBR) 1 and CBR3, which surround the C2 domain calcium-binding sites, whose mutation dramatically decreased phospholipid binding in vitro without significantly affecting calcium binding. Mutation of a residue that binds calcium ions (D43N) also eliminated phospholipid binding. The same mutations that prevent phospholipid binding of the isolated C2 domain in vitro abolished the calcium-dependent translocation of cPLA 2 to internal membranes in vivo, suggesting that the membrane targeting is driven largely by direct interactions with the phospholipid bilayer. Using fluorescence quenching by spin-labeled phospholipids for a series of mutants containing a single tryptophan residue at various positions in the cPLA 2 C2 domain, we show that two of the calcium-binding loops, CBR1 and CBR3, penetrate in a calcium-dependent manner into the hydrophobic core of the phospholipid bilayer, establishing an anchor for docking the domain onto the membrane.Cytosolic phospholipase A 2 (cPLA 2 ) 1 is an 85-kDa protein that hydrolyzes phospholipids containing arachidonate at the sn-2 position (Refs. 1 and 2; reviewed in Refs. 3-6). This enzyme has no sequence homology to any other phospholipase A 2 and is capable of functioning in receptor-regulated, agonistinduced arachidonic acid release (7,8). Recent work with mice lacking a gene for cPLA 2 has demonstrated that cPLA 2 has a critical, nonredundant role in the production of eicosanoids and platelet-activating factor in the process of inflammation, anaphylaxis, and reproduction (9, 10). The activity of cPLA 2 is regulated by calcium; however, the role of calcium is to promote membrane binding rather than participating in catalysis directly. An increase in intracellular calcium triggered by calcium ionophores or agonists such as histamine or IgE/antigen causes the translocation of cPLA 2 from the cytosol to the nuclear membrane and endoplasmic reticulum (11)(12)(13)(14), where it co-localizes with other enzymes involved in eicosanoid metabolism, such as prostaglandin-endoperoxide synthase-1 and -2, 5-lipoxygenase, and 5-lipoxygenase-activating protein (11, 15). The activity of cPLA 2 is also regulated via phosphorylation of the enzyme, and for at least some agonists, phosphorylation of cPLA 2 is a requisite step in its activation (4, 5). Detailed studies of the role of calcium and phosphorylation in arachidonic acid release have shown that a sustained increase in intracellular calcium is sufficient to induce arachidonic acid release, whereas either sustained phosphorylation of cPLA 2 or a transient ...

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“…Interestingly, the novel C2 domain factor has only 7 β strands (Fig. 2) in comparison with 8 stranded β-sandwich in other C2 domains of C2-domain superfamily proteins, such as PLC-δ, SynC2A, cPLA2, PKC-δ and PKC-β (Sutton et al, 1995;Essen et al, 1996;Perisic et al, 1998;Sutton and Sprang, 1998;Pappa et al, 1998). Also, the four putative proteins in zebrafish (GenBank accession no.…”

Section: Screening Cloning and Characterization Of A Novel C2 Domainmentioning

confidence: 99%

Identification of a novel C2 domain factor in ovaries of orange-spotted grouper (Epinephelus coioides)

Ji¹,

Zhou²,

Wang³

et al. 2006

Comparative Biochemistry and Physiology Part B: Biochemistry an

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Crystal structure of the C2 domain from protein kinase C-δ

Cited by 111 publications

References 41 publications

Structural Characterization of the C2 Domains of Classical Isozymes of Protein Kinase C and Novel Protein Kinase Cε by using Infrared Spectroscopy

Structural Characterization of the C2 Domains of Classical Isozymes of Protein Kinase C and Novel Protein Kinase Cε by using Infrared Spectroscopy

Mapping the Phospholipid-binding Surface and Translocation Determinants of the C2 Domain from Cytosolic Phospholipase A2

Identification of a novel C2 domain factor in ovaries of orange-spotted grouper (Epinephelus coioides)

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