TY  - JOUR
AU  - Margetić, Aleksandra
AU  - Nannemann, David
AU  - Meiler, Jens
AU  - Huster, Daniel
AU  - Theisgen, Stephan
PY  - 2014
UR  - https://cer.ihtm.bg.ac.rs/handle/123456789/1441
AB  - GCAPs are neuronal Ca2+-sensors playing a central role in light adaptation. GCAPs are N-terminally myristoylated. membrane-associated proteins. Although, the myristoylation of GCAPs plays an important role in light adaptation its structural and physiological roles are not yet clearly understood. The crystal-structure of GCAP-1 shows the myristoyl moiety inside the hydrophobic core of the protein, stabilizing the protein structure; but H-2-solid-state NMR investigations on the deuterated myristoyl moiety of GCAP-2 in the presence of liposomes showed that it is inserted into the lipid bilayer. In this study, we address the question of the localization of the myristoyl group of Ca2+-bound GCAP-2, and the influence of CHAPS-, DPC-micelles and DMPC/DHPC-bicelles on the structure, and on the localization of the myristoyl group, of GCAP-2 by solution-state NMR. We also carried out the backbone assignment. Characteristic chemical shift differences have been observed between the myristoylated and the non-myristoylated forms of the protein. Our results support the view that in the absence of membrane forming substances the myristoyl moiety is buried inside a hydrophobic pocket of GCAP-2 similar to the crystal structure of GCAP-1. Addition of CHAPS-micelles and DMPC/DHPC-bicelles cause specific structural changes localized in and around the myristoyl binding pocket. We interpret these changes as an indication for the extrusion of the myristoyl moiety from its binding pocket and its insertion into the hydrophobic interior of the membrane mimic. On the basis of the backbone chemical shifts, we propose a structural model of myristoylated GCAP-2 in the presence of Ca2+ and membrane mimetics.
PB  - Elsevier
T2  - Biochimica et Biophysica Acta-Biomembranes
T1  - Guanylate Cyclase-Activating Protein-2 Undergoes Structural Changes upon Binding to Detergent Micelles and Bicelles
VL  - 1838
IS  - 11
SP  - 2767
EP  - 2777
DO  - 10.1016/j.bbamem.2014.07.012
ER  - 

@article{
author = "Margetić, Aleksandra and Nannemann, David and Meiler, Jens and Huster, Daniel and Theisgen, Stephan",
year = "2014",
abstract = "GCAPs are neuronal Ca2+-sensors playing a central role in light adaptation. GCAPs are N-terminally myristoylated. membrane-associated proteins. Although, the myristoylation of GCAPs plays an important role in light adaptation its structural and physiological roles are not yet clearly understood. The crystal-structure of GCAP-1 shows the myristoyl moiety inside the hydrophobic core of the protein, stabilizing the protein structure; but H-2-solid-state NMR investigations on the deuterated myristoyl moiety of GCAP-2 in the presence of liposomes showed that it is inserted into the lipid bilayer. In this study, we address the question of the localization of the myristoyl group of Ca2+-bound GCAP-2, and the influence of CHAPS-, DPC-micelles and DMPC/DHPC-bicelles on the structure, and on the localization of the myristoyl group, of GCAP-2 by solution-state NMR. We also carried out the backbone assignment. Characteristic chemical shift differences have been observed between the myristoylated and the non-myristoylated forms of the protein. Our results support the view that in the absence of membrane forming substances the myristoyl moiety is buried inside a hydrophobic pocket of GCAP-2 similar to the crystal structure of GCAP-1. Addition of CHAPS-micelles and DMPC/DHPC-bicelles cause specific structural changes localized in and around the myristoyl binding pocket. We interpret these changes as an indication for the extrusion of the myristoyl moiety from its binding pocket and its insertion into the hydrophobic interior of the membrane mimic. On the basis of the backbone chemical shifts, we propose a structural model of myristoylated GCAP-2 in the presence of Ca2+ and membrane mimetics.",
publisher = "Elsevier",
journal = "Biochimica et Biophysica Acta-Biomembranes",
title = "Guanylate Cyclase-Activating Protein-2 Undergoes Structural Changes upon Binding to Detergent Micelles and Bicelles",
volume = "1838",
number = "11",
pages = "2767-2777",
doi = "10.1016/j.bbamem.2014.07.012"
}

Margetić, A., Nannemann, D., Meiler, J., Huster, D.,& Theisgen, S.. (2014). Guanylate Cyclase-Activating Protein-2 Undergoes Structural Changes upon Binding to Detergent Micelles and Bicelles. in Biochimica et Biophysica Acta-Biomembranes
Elsevier., 1838(11), 2767-2777.
https://doi.org/10.1016/j.bbamem.2014.07.012

Margetić A, Nannemann D, Meiler J, Huster D, Theisgen S. Guanylate Cyclase-Activating Protein-2 Undergoes Structural Changes upon Binding to Detergent Micelles and Bicelles. in Biochimica et Biophysica Acta-Biomembranes. 2014;1838(11):2767-2777.
doi:10.1016/j.bbamem.2014.07.012 .

Margetić, Aleksandra, Nannemann, David, Meiler, Jens, Huster, Daniel, Theisgen, Stephan, "Guanylate Cyclase-Activating Protein-2 Undergoes Structural Changes upon Binding to Detergent Micelles and Bicelles" in Biochimica et Biophysica Acta-Biomembranes, 1838, no. 11 (2014):2767-2777,
https://doi.org/10.1016/j.bbamem.2014.07.012 . .