Identification of the binding domain for secretory phospholipases A2 on their M-type 180-kDa membrane receptor.

The rabbit muscle (M)-type receptor for secretory phospholipases A2 (sPLA2s) has a large extracellular domain of 1394 amino acids, composed of an N-terminal cysteine-rich domain, a fibronectin-like type II domain, and eight carbohydrate recognition domains (CRDs). It is thought to mediate some of the physiological effects of mammalian sPLA2s, including vascular smooth muscle contraction and cell proliferation, and is able to internalize sPLA2s. Here, we show by site-directed mutagenesis that OS1, a snake venom sPLA2, binds to the receptor via its CRDs and that deletion of CRD 5 completely abolishes the binding of sPLA2s. Moreover, a receptor lacking all CRDs but CRD 5 was still able to bind OS1 although with a lower affinity. Deletion of CRDs 4 and 6, surrounding the CRD 5, slightly reduced the affinity for OS1, thus suggesting that these CRDs are also involved in the binding of OS1. The M-type sPLA2 receptor and the macrophage mannose receptor are homologous and are predicted to share the same tertiary structure. p-Aminophenyl-alpha-D-mannopyranoside bovine serum albumin, a known ligand of the macrophage mannose receptor, binds to the M-type sPLA2 receptor essentially via CRDs 3-6.