Binding of moesin and ezrin to membranes containing phosphatidylinositol (4,5) bisphosphate: a comparative study of the affinity constants and conformational changes.
The plasma membrane-cytoskeleton interface is a dynamic structure participating in a variety of cellular events. Moesin and ezrin, proteins from the ezrin/radixin/moesin (ERM) family, provide a direct linkage between the cytoskeleton and the membrane via their interaction with phosphatidylinositol 4,5-bisphosphate (PIP(2)). PIP(2) binding is considered as a prerequisite step in ERM activation. The main objective of this work was to compare moesin and ezrin interaction with PIP(2)-containing membranes in terms of affinity and to analyze secondary structure modifications leading eventually to ERM activation. For this purpose, we used two types of biomimetic model membranes, large and giant unilamellar vesicles. The dissociation constant between moesin and PIP(2)-containing large unilamellar vesicles or PIP(2)-containing giant unilamellar vesicles was found to be very similar to that between ezrin and PIP(2)-containing large unilamellar vesicles or PIP(2)-containing giant unilamellar vesicles. In addition, both proteins were found to undergo conformational changes after binding to PIP(2)-containing large unilamellar vesicles. Changes were evidenced by an increased sensitivity to proteolysis, modifications in the fluorescence intensity of the probe attached to the C-terminus and in the proportion of secondary structure elements.
Grenoble Institute of Technology and CNRS, Grenoble Cedex, France.
Ofelia
O
Maniti
Nada
N
Khalifat
Kriti
K
Goggia
Fabien
F
Dalonneau
Christophe
C
Guérin
Laurent
L
Blanchoin
Laurence
L
Ramos
Catherine
C
Picart
D032701:Biomimetics; D003598:Cytoskeletal Proteins / Q000737:chemistry / Q000378:metabolism*; D008567:Membranes, Artificial; D008840:Microfilament Proteins / Q000737:chemistry / Q000378:metabolism*; D008958:Models, Molecular; D008968:Molecular Conformation; D019269:Phosphatidylinositol 4,5-Diphosphate / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D059748:Proteolysis; D017550:Spectroscopy, Fourier Transform Infrared; D000099149:Ezrin
D003598:Cytoskeletal Proteins; D008567:Membranes, Artificial; D008840:Microfilament Proteins; D019269:Phosphatidylinositol 4,5-Diphosphate; D000099149:Ezrin; C071058:moesin