Structural changes in profilin accompany its binding to phosphatidylinositol, 4,5-bisphosphate.
The effect on the structure of profilin of phosphatidylinositol 4,5-bisphosphate (PIP2) binding was probed by fluorescence and circular dichroism (CD) spectroscopy. Fluorescence of Trp3 and Trp31 of profilin at 292 nm showed a linear decrease in solution emission at 340 nm as PIP2/profilin was increased from 0 to 80:1, apparently due to a static quenching mechanism involving formation of a nonfluorescent PIP2/profilin complex. CD spectra revealed an increase of up to 3.3-fold in the molar ellpticity at 222 nm for profilin as it binds PIP2, as well as changes in the Cotton effect between 250 and 310 nm. These results are consistent with a possible increase in the alpha-helix content of profilin triggered by the binding of PIP2.
Raghunathan
V
V
Frick Chemical Laboratory, Princeton University, NJ 08544.
Mowery
P
P
Rozycki
M
M
Lindberg
U
U
Schutt
C
C
D000818:Animals; D002417:Cattle; D002942:Circular Dichroism; D003285:Contractile Proteins; D008840:Microfilament Proteins / Q000378:metabolism*; D019269:Phosphatidylinositol 4,5-Diphosphate; D010716:Phosphatidylinositols / Q000378:metabolism*; D051304:Profilins; D011487:Protein Conformation; D013050:Spectrometry, Fluorescence
D003285:Contractile Proteins; D008840:Microfilament Proteins; D019269:Phosphatidylinositol 4,5-Diphosphate; D010716:Phosphatidylinositols; D051304:Profilins