Retinoic Acid Binds to the C2-Domain of Protein Kinase Cα

• Published in: Biochemistry (Easton) Vol. 42; no. 29; pp. 8774 - 8779
• Main Authors: Ochoa, Wendy F, Torrecillas, Alejandro, Fita, Ignacio, Verdaguer, Nuria, Corbalán-García, Senena, Gomez-Fernandez, Juan C
• Format: Journal Article
• Language: English
• Published: American Chemical Society 29.07.2003
• ISSN: 0006-2960; 1520-4995
• DOI: 10.1021/bi034713g

Protein kinase Cα (PKCα) is a key enzyme regulating the physiology of cells and their growth, differentiation, and apoptosis. PKC activity is known to be modulated by all-trans retinoic acid (atRA), although neither the action mechanism nor even the possible binding to PKCs has been established. Crystals of the C2-domain of PKCα, a regulatory module in the protein that binds Ca2+ and acidic phospholipids, have now been obtained by cocrystallization with atRA. The crystal structure, refined at 2.0 Å resolution, shows that RA binds to the C2-domain in two locations coincident with the two binding sites previously reported for acidic phospholipids. The first binding site corresponds to the Ca2+-binding pocket, where Ca2+ ions mediate the interactions of atRA with the protein, as they do with acidic phospholipids. The second binding site corresponds to the conserved lysine-rich cluster localized in β-strands three and four. These observations are strongly supported by [3H]-atRA-binding experiments combined with site-directed mutagenesis. Wild-type C2-domain binds 2 mol of atRA per mol of protein, while the rate reduces to one in the case of C2-domain variants, in which mutations affect either Ca2+ coordination or the integrity of the lysine-rich cluster site. Competition between atRA and acidic phospholipids to bind to PKC is a possible mechanism for modulating PKCα activity.