Cyclase-Associated Protein 1 (CAP1) is a prenyl-binding partner of Rap1 GTPase

XUEFENG ZHANGA; SHUFEN CAO; GUILLERMO BARILA; MARTIN M EDREIRA ; MAMTA WANKHEDE ; NYLA NAIM; MATTHIAS BUCK; DANIEL L ALTSCHULER

JOURNAL OF BIOLOGICAL CHEMISTRY

AMER SOC BIOCHEMISTRY MOLECULAR BIOLOGY INC

Lugar: Bethesda, Maryland; Año: 2018 vol. 293 p. 7659 - 7673

0021-9258

Rap1 proteins are members of the Ras subfamily of small GTPases involved in many biological responses, including adhesion, cell proliferation, and differentiation. Like all small GTPases, they work as molecular allosteric units that are active in signaling only when associated with the proper membrane compartment. Prenylation, occurring in the cytosol, is an enzymatic posttranslational event that anchors small GTPases at the membrane, and prenyl-binding proteins are needed to mask the cytoplasm-exposed lipid during transit to the target membrane. However, several of these proteins still await discovery. In this study, we report that cyclase-associated protein 1 (CAP1) binds Rap1. We found that this binding is GTP-independent, does not involve Rap1?s effector domain, and is fully contained in its C-terminal hypervariable region (HVR). Furthermore, Rap1 prenylation was required for high-affinity interactions with CAP1 in a geranylgeranyl-specific manner. The prenyl binding specifically involved CAP1?s C-terminal hydrophobic β-sheet domain. We present a combination of experimental and computational approaches, yielding a model whereby the high-affinity binding between Rap1 and CAP1 involves electrostatic and nonpolar sidechain interactions between Rap1?s HVR residues, lipid and CAP1 β-sheet domain. The binding was stabilized by the lipid insertion into the elliptical β-barrel whose interior was occupied by nonpolar sidechains. This model was reminiscent of the recently solved structure of the PDEδ?K-Ras complex; accordingly, disruptors of this complex, e.g. Deltarasin, blocked the Rap1?CAP1 interaction. These findings indicate that CAP1 is a geranylgeranyl-binding partner of Rap1.