TY - JOUR
T1 - Characterization of the structure and function of W → F WW domain variants
T2 - Identification of a natively unfolded protein that folds upon ligand binding
AU - Koepf, Edward K.
AU - Petrassi, H. Michael
AU - Ratnaswamy, Gayathri
AU - Huff, Mary E.
AU - Sudol, Marius
AU - Kelly, Jeffery W.
PY - 1999/10/26
Y1 - 1999/10/26
N2 - The WW domain adopts a compact, three-stranded, antiparallel β-sheet structure that mediates protein-protein interactions by binding to xPPxY- based protein ligands, such as the PY-ligand (EYPPYPPPPYPSG) derived from p53 binding protein-2. The conserved Trp residues, after which this domain was named, were replaced with Phe so their importance in structural integrity and for ligand binding could be evaluated. A biophysical approach was employed to compare the W17F, W39F, and W17F/W39F WW domains to the wild-type protein. The data demonstrate that replacement of Trp39 with Phe (W39F) does not disrupt the structure of the WW domain variant, but does abolish ligand binding. In contrast, the W17F WW domain variant is largely if not completely unfolded; however, this variant undergoes a PY-ligand induced disorder to order (folding) transition. The dissociation constant for the W17F WW domain- PY-ligand interaction is 15.1 ± 1.2 μM, only slightly higher than that observed for the wild-type WW domain interaction (5.9 ± 0.33 μM). The W17F WW domain is a natively unfolded protein which adopts a native conformation upon PY-ligand binding.
AB - The WW domain adopts a compact, three-stranded, antiparallel β-sheet structure that mediates protein-protein interactions by binding to xPPxY- based protein ligands, such as the PY-ligand (EYPPYPPPPYPSG) derived from p53 binding protein-2. The conserved Trp residues, after which this domain was named, were replaced with Phe so their importance in structural integrity and for ligand binding could be evaluated. A biophysical approach was employed to compare the W17F, W39F, and W17F/W39F WW domains to the wild-type protein. The data demonstrate that replacement of Trp39 with Phe (W39F) does not disrupt the structure of the WW domain variant, but does abolish ligand binding. In contrast, the W17F WW domain variant is largely if not completely unfolded; however, this variant undergoes a PY-ligand induced disorder to order (folding) transition. The dissociation constant for the W17F WW domain- PY-ligand interaction is 15.1 ± 1.2 μM, only slightly higher than that observed for the wild-type WW domain interaction (5.9 ± 0.33 μM). The W17F WW domain is a natively unfolded protein which adopts a native conformation upon PY-ligand binding.
UR - http://www.scopus.com/inward/record.url?scp=0033607151&partnerID=8YFLogxK
U2 - 10.1021/bi991105l
DO - 10.1021/bi991105l
M3 - Article
C2 - 10572009
AN - SCOPUS:0033607151
SN - 0006-2960
VL - 38
SP - 14338
EP - 14351
JO - Biochemistry
JF - Biochemistry
IS - 43
ER -