Binding of a Monoclonal Antibody to the Phospholamban Cytoplasmic Domain Interferes with the Channel Activity of Phospholamban Reconstituted in a Tethered Bilayer Lipid Membrane
journal contributionposted on 2014-09-02, 00:00 authored by Serena Smeazzetto, Alessio Sacconi, Adrian L. Schwan, Giancarlo Margheri, Francesco Tadini-Buoninsegni
Phospholamban (PLN), a membrane protein present in the sarcoplasmic reticulum of cardiac myocytes, is a crucial regulator of cardiac function. It is known that PLN appears as a monomer and as a pentamer. However, the role of the PLN pentamer and its ability to generate an ion channel are a matter of debate. To address this issue we employed an experimental approach that combines electrochemical impedance spectroscopy and surface plasmon resonance measurements. In particular, we investigated the channel activity of wild-type PLN reconstituted in a tethered bilayer lipid membrane (tBLM) on a gold surface. Our results indicate that reconstituted PLN can generate ion-conducting channels in a tBLM. Experiments with a PLN monoclonal antibody support an oriented incorporation of PLN in the tBLM. We show that the binding of the antibody to the PLN cytoplasmic domain interferes with PLN channel activity.
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Monoclonal Antibodysurface plasmon resonance measurementsPLN channel activitymembrane proteinbilayer lipid membraneantibody supportPLN pentamerelectrochemical impedance spectroscopyreconstituted PLNchannel activityPhospholamban Reconstitutedion channelgold surfacePLN cytoplasmic domainChannel Activitysarcoplasmic reticulumPhospholamban Cytoplasmic Domain InterferesTethered Bilayer Lipid MembranePhospholamban