P. Garcia et al., THE PLECKSTRIN HOMOLOGY DOMAIN OF PHOSPHOLIPASE C-DELTA(1) BINDS WITHHIGH-AFFINITY TO PHOSPHATIDYLINOSITOL 4,5-BISPHOSPHATE IN BILAYER-MEMBRANES, Biochemistry, 34(49), 1995, pp. 16228-16234
The pleckstrin homology (PH) domain of phospholipase C-delta(1) (PLC-d
elta(1)) binds to phosphatidylinositol 4,5-bisphosphate (PI(4,5)P-2) i
n phospholipid membranes with an affinity (K-a similar to 10(6) M(-1))
and specificity comparable to those of the native enzyme. PLC-delta(1
) and its PH domain also bind inositol 1,4,5-trisphosphate, the polar
head group of PI(4,5)P-2, with comparable affinity and approximately 1
:1 stoichiometry. A peptide corresponding to amino acids 30-43 of the
PLC-delta(1) PH domain contains several basic residues predicted to bi
nd PI(4,5)P-2, but binds weakly and with little specificity for PI(4,5
)P-2; hence the tertiary structure of the isolated PH domain is requir
ed for high affinity PI(4,5)P-2 binding. Our PI(4,5)P-2 binding result
s support the hypothesis that the intact PH domain, serving as' a spec
ific tether, directs PLC-delta(1) to membranes enriched in PI(4,5)P-2
and permits the active site, located elsewhere in the protein, to hydr
olyze multiple substrate molecules before this enzyme dissociates from
the membrane surface.