Crystal structure of a calcium-phospholipid binding domain from cytosolic phospholipase A2.
Cytosolic phospholipase A2 (cPLA2) is a calcium-sensitive 85-kDa enzyme that hydrolyzes arachidonic acid-containing membrane phospholipids to initiate the biosynthesis of eicosanoids and platelet-activating factor, potent inflammatory mediators. The calcium-dependent activation of the enzyme is mediated by an N-terminal C2 domain, which is responsible for calcium-dependent translocation of the enzyme to membranes and that enables the intact enzyme to hydrolyze membrane-resident substrates. The 2.4-A x-ray crystal structure of this C2 domain was solved by multiple isomorphous replacement and reveals a beta-sandwich with the same topology as the C2 domain from phosphoinositide-specific phospholipase C delta 1. Two clusters of exposed hydrophobic residues surround two adjacent calcium binding sites. This region, along with an adjoining strip of basic residues, appear to constitute the membrane binding motif. The structure provides a striking insight into the relative importance of hydrophobic and electrostatic components of membrane binding for cPLA2. Although hydrophobic interactions predominate for cPLA2, for other C2 domains such as in "conventional" protein kinase C and synaptotagmins, electrostatic forces prevail.
Perisic
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Medical Research Council Laboratory of Molecular Biology, Medical Research Council Centre, Cambridge, United Kingdom.
Fong
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Lynch
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Bycroft
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Williams
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D000595:Amino Acid Sequence; D001665:Binding Sites; D002118:Calcium / Q000378:metabolism*; D003001:Cloning, Molecular; D018360:Crystallography, X-Ray; D003600:Cytosol / Q000201:enzymology; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism; D008956:Models, Chemical; D008958:Models, Molecular; D008969:Molecular Sequence Data; D054803:Phospholipase C delta; D010741:Phospholipases A / Q000737:chemistry*; D054467:Phospholipases A2; D010743:Phospholipids / Q000378:metabolism*; D017433:Protein Structure, Secondary; D010738:Type C Phospholipases / Q000378:metabolism
D007527:Isoenzymes; D010743:Phospholipids; D010741:Phospholipases A; D054467:Phospholipases A2; D010738:Type C Phospholipases; C518024:PLCD1 protein, human; D054803:Phospholipase C delta; D002118:Calcium