Evidence for direct interaction between actin and the cystic fibrosis transmembrane conductance regulator.
Previous studies have demonstrated that actin filament organization controls the cystic fibrosis transmembrane conductance regulator (CFTR) ion channel function. The precise molecular nature of the interaction between actin and CFTR, however, remains largely unknown. In this report, interactions between actin and purified human epithelial CFTR were directly assessed by reconstitution of the channel protein in a lipid bilayer system and by atomic force microscopy (AFM). CFTR-containing liposomes in solution were deposited on freshly cleaved mica and imaging was performed in tapping-mode AFM. CFTR function was also determined in identical preparations. Images of single CFTR molecules were obtained, and addition of monomeric actin below its critical concentration showed the formation of actin filaments associated with CFTR. The data indicate a direct interaction between actin and CFTR exists, which may explain the regulatory role of the cytoskeleton in ion channel function. This was confirmed by functional studies of CFTR single-channel currents, which were regulated by addition of various conformations of actin. The present study indicates that CFTR may directly bind actin and that this interaction helps affect the functional properties of this channel protein.
Chasan
Bernard
B
Physics Department, Boston University, MA, USA.
Geisse
Nicholas A
NA
Pedatella
Kate
K
Wooster
David G
DG
Teintze
Martin
M
Carattino
Marcelo D
MD
Goldmann
Wolfgang H
WH
Cantiello
Horacio F
HF
D000199:Actins / Q000378:metabolism*; D000818:Animals; D002460:Cell Line; D000242:Cyclic AMP / Q000378:metabolism; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000378:metabolism*; D003599:Cytoskeleton / Q000378:metabolism; D004594:Electrophysiology; D006801:Humans; D007313:Insecta; D007477:Ions; D008051:Lipid Bilayers / Q000378:metabolism; D018625:Microscopy, Atomic Force; D011485:Protein Binding; D011721:Pyrenes / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D014965:X-Rays
D000199:Actins; C505032:CFTR protein, human; D007477:Ions; D008051:Lipid Bilayers; D011721:Pyrenes; D011994:Recombinant Proteins; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D000242:Cyclic AMP