Measurement of membrane binding between recoverin, a calcium-myristoyl switch protein, and lipid bilayers by AFM-based force spectroscopy.
Myristoyl switch is a feature of several peripheral membrane proteins involved in signal transduction pathways. This unique molecular property is best illustrated by the "Ca(2+)-myristoyl switch" of recoverin, which is a Ca(2+)-binding protein present in retinal rod cells of vertebrates. In this transduction pathway, the Ca(2+)-myristoyl switch acts as a calcium sensor involved in cell recovery from photoactivation. Ca(2+) binding by recoverin induces the extrusion of its myristoyl group to the solvent, which leads to its translocation from cytosol to rod disk membranes. Force spectroscopy, based on atomic force microscope (AFM) technology, was used to determine the extent of membrane binding of recoverin in the absence and presence of calcium, and to quantify this force of binding. An adhesion force of 48 +/- 5 pN was measured between recoverin and supported phospholipid bilayers in the presence of Ca(2+). However, no binding was observed in the absence of Ca(2+). Experiments with nonmyristoylated recoverin confirmed these observations. Our results are consistent with previously measured extraction forces of lipids from membranes.
Desmeules
Philippe
P
Département de Chimie-Biologie, Université du Québec à Trois-Rivières, Trois-Rivières, Québec G9A 5H7, Canada.
Grandbois
Michel
M
Bondarenko
Vladimir A
VA
Yamazaki
Akio
A
Salesse
Christian
C
D015060:1,2-Dipalmitoylphosphatidylcholine / Q000737:chemistry; D000818:Animals; D055592:Biophysical Phenomena; D001703:Biophysics; D002118:Calcium / Q000378:metabolism; D002135:Calcium-Binding Proteins / Q000737:chemistry* / Q000378:metabolism*; D005136:Eye Proteins / Q000737:chemistry / Q000378:metabolism; D051597:Hippocalcin; D066298:In Vitro Techniques; D008051:Lipid Bilayers / Q000737:chemistry* / Q000378:metabolism*; D008074:Lipoproteins; D018625:Microscopy, Atomic Force; D008958:Models, Molecular; D019814:Myristic Acid / Q000737:chemistry; D009419:Nerve Tissue Proteins; D011485:Protein Binding; D011487:Protein Conformation; D011994:Recombinant Proteins / Q000737:chemistry / Q000378:metabolism; D051603:Recoverin; D015398:Signal Transduction
D002135:Calcium-Binding Proteins; D005136:Eye Proteins; D008051:Lipid Bilayers; D008074:Lipoproteins; D009419:Nerve Tissue Proteins; D011994:Recombinant Proteins; D019814:Myristic Acid; D051603:Recoverin; D051597:Hippocalcin; D015060:1,2-Dipalmitoylphosphatidylcholine; D002118:Calcium