TY - JOUR
T1 - Structure and phosphatidylinositol-(3,4)-bisphosphate binding of the C-terminal PH domain of human pleckstrin
AU - Edlich, Christian
AU - Stier, Gunter
AU - Simon, Bernd
AU - Sattler, Michael
AU - Muhle-Goll, Claudia
N1 - Funding Information:
We acknowledge the support from the DFG and the Volkswagen Stiftung. C.M.-G. is an Emmy Noether-fellow of the DFG. We would like to thank Fabien Bonneau and Massimiliano Mazza, EMBL, for support with protein lipid overlay assays; Carsten Schultz and Andreas Schleifenbaum for fruitful discussions; and to Professor D. Alessi for providing the TAPP1 clone.
PY - 2005/2
Y1 - 2005/2
N2 - Pleckstrin is the major target of protein kinase C (PKC) in blood platelets. Its phosphorylation triggers responses that ultimately lead to platelet activation and blood clot formation. Pleckstrin consists of three domains: a pleckstrin homology (PH) domain at both termini and a central DEP (Dishevelled, Egl-1, Pleckstrin) domain. Here, we report the solution nuclear magnetic resonance (NMR) structure of the C-terminal PH domain (C-PH) of human pleckstrin-1. We show that this PH domain binds phosphatidylinositol-3,4- bisphosphate (PtdIns(3,4)P2) with high specificity in protein lipid overlay assays. Using NMR titration experiments and mutational analysis, residues involved in binding to PtdIns(3,4)P2 are identified. The binding site is formed by a patch of basic residues from the β1 and β2 strands and the β1-β2 loop. Since PtdIns(3,4)P2 is an important signaling molecule in platelets, our data suggest a C-PH dependent regulation of pleckstrin function in response to PtdIns(3,4)P2.
AB - Pleckstrin is the major target of protein kinase C (PKC) in blood platelets. Its phosphorylation triggers responses that ultimately lead to platelet activation and blood clot formation. Pleckstrin consists of three domains: a pleckstrin homology (PH) domain at both termini and a central DEP (Dishevelled, Egl-1, Pleckstrin) domain. Here, we report the solution nuclear magnetic resonance (NMR) structure of the C-terminal PH domain (C-PH) of human pleckstrin-1. We show that this PH domain binds phosphatidylinositol-3,4- bisphosphate (PtdIns(3,4)P2) with high specificity in protein lipid overlay assays. Using NMR titration experiments and mutational analysis, residues involved in binding to PtdIns(3,4)P2 are identified. The binding site is formed by a patch of basic residues from the β1 and β2 strands and the β1-β2 loop. Since PtdIns(3,4)P2 is an important signaling molecule in platelets, our data suggest a C-PH dependent regulation of pleckstrin function in response to PtdIns(3,4)P2.
UR - http://www.scopus.com/inward/record.url?scp=13844253933&partnerID=8YFLogxK
U2 - 10.1016/j.str.2004.11.012
DO - 10.1016/j.str.2004.11.012
M3 - Article
C2 - 15698571
AN - SCOPUS:13844253933
SN - 0969-2126
VL - 13
SP - 277
EP - 286
JO - Structure
JF - Structure
IS - 2
ER -