resumo
Clathrin assembly lymphoid myeloid leukemia protein (CALM) is involved in the formation of clathrin-mediated endocytic coats by virtue of binding many proteins involved in the process, including clathrin itself and AP2 cargo adaptor complex. CALM is able to specifically recognize the inner leaflet of the plasma membrane by binding the membrane’s phosphatidylinositol 4,5-bisphosphate (PtdIns(4,5)P2). Here, a biophysical approach, primarily using neutron and X-ray scattering and solid-state NMR experiments, was exploited to investigate CALM interaction with PtdIns(4,5)P2-presenting model membranes. The presented experimental data reveal how the CALM folded domain is partly accommodated within the lipid membrane, directly interacting with PtdIns(4,5)P2 phosphates. Moreover, these data suggest that CALM’s amphiphilic N-terminal helix buries into the membrane, not only stabilising the protein docking to the membrane but also providing a mechanism to induce membrane curvature.
autores
Andreas Santamaria, Daniel Pereira, Nisha Pawar, Bernard T. Kelly, Javier Carrascosa-Tejedor, Mariana Sardo, Luís Mafra, Giovanna Fragneto, David J. Owen, Ildefonso Marín-Montesinos, Eduardo Guzmán, Nathan R. Zaccai, Armando Maestro
nossos autores
Daniel António Cunha Pereira
Estudante de doutoramento
Ildefonso Marin Montesinos
Investigador Júnior
Luís Mafra
Investigador Principal
Mariana Coutinho Sardo
Investigador AuxiliarProjectos
CICECO - Aveiro Institute of Materials (UIDP/50011/2020)
Associated Laboratory CICECO-Aveiro Institute of Materials (LA/P/0006/2020)
agradecimentos
The authors thank both the Institut Laue-Langevin (DOI:ILL-DATA.8-02-829) and the European Synchrotron Radiation Facility ESRF (DOI:10.15151/ESRF-ES-883953841) for the allocation of beamtime and the Partnership for Soft Condensed Matter (PSCM) for the lab support. A.M. acknowledges the financial support from MCIN/AEI/10.13039/501100011033 under grant PID2021-129054NA-I00, and from the Department of Education of the Basque Government under grant PIBA_2023_1_0054 and from the IKUR Strategy under the collaboration agreement between Ikerbasque Foundation and Materials Physics Center. E.G. acknowledge the financial support MCIN/AEI/10.13039/501100011033 and UCM under grants PID2019-106557GB-C21 and PR12/24-31566 (Ayudas para la financiación de proyectos de investigación UCM 2023). NZ and DJO acknowledge the support of the Wellcome Trust grant to DO (207455/Z/17/Z). This work was also developed within the scope of project CICECO-Aveiro Institute of Materials, UIDB/50011/ 2020, UIDP/50011/2020 & LA/P/0006/2020, financed by national funds through the FCT/MEC (PIDDAC). The NMR spectrometers are part of the National NMR Network (PTNMR) and are partially supported by Infrastructure Project 022161 (cofinanced by FEDER through COMPETE 2020, POCI and PORL and FCT through PIDDAC). FCT is also acknowledged by D. P. for a Ph.D. Studentship UI/BD/151048/2021 (DOI: 10.54499/UI/BD/151048/2021). I.M.-M. acknowledges the EMBO organisation for the EMBO Fellowship 8740.