Crystal structure of the phosphatidylinositol 3,4-bisphosphate-binding pleckstrin homology (PH) domain of tandem PH-domain-containing protein 1 (TAPP1): molecular basis of lipid specificity

摘要

pPhosphatidylinositol 3,4,5-trisphosphate [PtdIns(3,4,5)iP/isub3/sub] and its immediate breakdown product PtdIns(3,4)iP/isub2/sub function as second messengers in growth factor- and insulin-induced signalling pathways. One of the ways that these 3-phosphoinositides are known to regulate downstream signalling events is by attracting proteins that possess specific PtdIns-binding pleckstrin homology (PH) domains to the plasma membrane. Many of these proteins, such as protein kinase B, phosphoinositide-dependent kinase 1 and the bd/bual ba/bdaptor for bp/bhosphotyrosine and 3-bp/bhosphoinositides (DAPP1) interact with both PtdIns(3,4,5)iP/isub3/sub and PtdIns(3,4)iP/isub2/sub with similar affinity. Recently, a new PH-domain-containing protein, termed bta/bndem bP/bH-domain-containing bp/brotein (TAPP) 1, was described which is the first protein reported to bind PtdIns(3,4)iP/isub2/sub specifically. Here we describe the crystal structure of the PtdIns(3,4)iP/isub2/sub-binding PH domain of TAPP1 at 1.4 ? (1 ? = 0.1 nm) resolution in complex with an ordered citrate molecule. The structure is similar to the known structure of the PH domain of DAPP1 around the D-3 and D-4 inositol-phosphate-binding sites. However, a glycine residue adjacent to the D-5 inositol-phosphate-binding site in DAPP1 is substituted for a larger alanine residue in TAPP1, which also induces a conformational change in the neighbouring residues. We show that mutation of this glycine to alanine in DAPP1 converts DAPP1 into a TAPP1-like PH domain that only interacts with PtdIns(3,4)iP/isub2/sub, whereas the alanine to glycine mutation in TAPP1 permits the TAPP1 PH domain to interact with PtdIns(3,4,5)iP/isub3/sub./p