Nanomechanical properties of composite protein networks of erythroid membranes at lipid surfaces

Erythrocyte membranes have been particularly useful as a model for studies of membrane structure and mechanics. Native erythroid membranes can be electroformed as giant unilamellar vesicles (eGUVs). In the presence of ATP, the erythroid membrane proteins of eGUVs rearrange into protein networks at the microscale. Here, we present a detailed nanomechanical study of individual protein microfilaments forming the protein networks of eGUVs when spread on supporting surfaces. Using Peak Force tapping Atomic Force Microscopy (PF-AFM) in liquid environment we have obtained the mechanical maps of the composite lipid-protein networks supported on solid surface. In the absence of ATP, the protein pool was characterized by a Young’s Modulus Epool ≈ 5–15 MPa whereas the complex filaments were found softer after protein supramolecular rearrangement; Efil ≈ 0.4 MPa. The observed protein softening and reassembling could be relevant for understanding the mechanisms of cytoskeleton reorganization found in pathological erythrocytes or erythrocytes that are affected by biological agents.This work was supported by the ERC Starting Grant “mitochon” (ERC-StG-2013-883188) and “Programa Ramon y Cajal” (RYC-2013-12609) from the Spanish Ministry of Economy MINECO (I. L-M.), the ERC Starting Grant “NANOFORCELLS” (ERC-StG-2011-278860) (M. E., A. C-M., M. C.), the project FORCE-for-FUTURE (CSD2010-00024) (A. S. P.) and FIS2012-35723 from MINECO (F.M.).Peer reviewe