Comparison of VILIP-1 and VILIP-3 binding to phospholipid monolayers.
The neuronal calcium sensor proteins Visinin-like Proteins 1 (VILIP-1) and 3 (VILIP-3) are effectors of guanylyl cyclase and acetyl choline receptors, and transduce calcium signals in the brain. The "calcium-myristoyl" switch, which involves a post-translationally added myristoyl moiety and calcium binding, is thought to regulate their membrane binding capacity and therefore, play a critical role in their mechanism of action. In the present study, we investigated the effect of membrane composition and solvent conditions on the membrane binding mechanisms of both VILIPs using lipid monolayers at the air/buffer interface. Results based on comparison of the adsorption kinetics of the myristoylated and non-myristoylated proteins confirm the pivotal role of calcium and the exposed myristol moiety for sustaining the membrane-bound state of both VILIPs. However, we also observed binding of both VILIP proteins in the absence of calcium and/or myristoyl conjugation. We propose a two-stage membrane binding mechanism for VILIP-1 and VILIP-3 whereby the proteins are initially attracted to the membrane surface by electrostatic interactions and possibly by specific interactions with highly negatively charged lipids head groups. The extrusion of the conjugated myristoyl group, and the subsequent anchoring in the membrane constitutes the second stage of the binding mechanism, and ensures the sustained membrane-bound form of these proteins.
Institut de Chimie et Biochimie Moléculaires et Supramoléculaires, Université Lyon 1, University of Lyon, ICBMS, CNRS UMR 5246, Bât. Curien, 43 bd du 11 Nov. 1918, F-69622 Villeurbanne cedex, France.
Samuel
S
Rebaud
Institut de Chimie et Biochimie Moléculaires et Supramoléculaires, Université Lyon 1, University of Lyon, ICBMS, CNRS UMR 5246, Bât. Curien, 43 bd du 11 Nov. 1918, F-69622 Villeurbanne cedex, France.
Anne
A
Simon
Structural Chemistry Program, Eskitis Institute, Griffith University, Brisbane, Queensland, Australia.
Conan K
CK
Wang
Structural Chemistry Program, Eskitis Institute, Griffith University, Brisbane, Queensland, Australia.
Lyndel
L
Mason
Institut de Chimie et Biochimie Moléculaires et Supramoléculaires, Université Lyon 1, University of Lyon, ICBMS, CNRS UMR 5246, Bât. Curien, 43 bd du 11 Nov. 1918, F-69622 Villeurbanne cedex, France.
Loïc
L
Blum
Structural Chemistry Program, Eskitis Institute, Griffith University, Brisbane, Queensland, Australia.
Andreas
A
Hofmann
Institut de Chimie et Biochimie Moléculaires et Supramoléculaires, Université Lyon 1, University of Lyon, ICBMS, CNRS UMR 5246, Bât. Curien, 43 bd du 11 Nov. 1918, F-69622 Villeurbanne cedex, France.
Agnès
A
Girard-Egrot
D002118:Calcium / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism*; D004926:Escherichia coli / Q000378:metabolism; D051596:Neurocalcin / Q000378:metabolism*; D010743:Phospholipids / Q000378:metabolism*; D011485:Protein Binding
C520235:HPCAL1 protein, human; D051596:Neurocalcin; D010743:Phospholipids; C496688:VSNL1 protein, human; D002118:Calcium