Ca2+ bridges the C2 membrane-binding domain of protein kinase Cα directly to phosphatidylserine

• Published in: The EMBO journal 1999-11, Vol.18 (22), p.6329-6338
• Main Authors: Verdaguer, Nuria, Corbalan-Garcia, Senena, Ochoa, Wendy F., Fita, Ignacio, Gómez-Fernández, Juan C.
• Format: Article
• Language: English
• Subjects: C2 domain; Ca2+ binding; phosphatidylserine; protein kinase C; X-ray structure
• ISSN: 0261-4189; 1460-2075
• DOI: 10.1093/emboj/18.22.6329

The C2 domain acts as a membrane‐targeting module in a diverse group of proteins including classical protein kinase Cs (PKCs), where it plays an essential role in activation via calcium‐dependent interactions with phosphatidylserine. The three‐dimensional structures of the Ca 2+ ‐bound forms of the PKCα‐C2 domain both in the absence and presence of 1,2‐dicaproyl‐ sn ‐phosphatidyl‐L‐serine have now been determined by X‐ray crystallography at 2.4 and 2.6 Å resolution, respectively. In the structure of the C2 ternary complex, the glycerophosphoserine moiety of the phospholipid adopts a quasi‐cyclic conformation, with the phosphoryl group directly coordinated to one of the Ca 2+ ions. Specific recognition of the phosphatidylserine is reinforced by additional hydrogen bonds and hydrophobic interactions with protein residues in the vicinity of the Ca 2+ binding region. The central feature of the PKCα‐C2 domain structure is an eight‐stranded, anti‐parallel β‐barrel with a molecular topology and organization of the Ca 2+ binding region closely related to that found in PKCβ‐C2, although only two Ca 2+ ions have been located bound to the PKCα‐C2 domain. The structural information provided by these results suggests a membrane binding mechanism of the PKCα‐C2 domain in which calcium ions directly mediate the phosphatidylserine recognition while the calcium binding region 3 might penetrate into the phospholipid bilayer.