Abstract
The solid supported membrane (SSM) represents a convenient model system for a biological membrane with the advantage of being mechanically so stable that solutions can be rapidly exchanged at the surface. The SSM consists of a hybrid alkanethiol–phospholipid bilayer supported by a gold electrode. Proteoliposomes, membrane vesicles, or membrane fragments containing the transport protein of interest are adsorbed on the SSM surface and are subjected to a rapid substrate concentration jump. The substrate concentration jump activates the protein and the charge displacement concomitant with its transport activity is recorded as a current transient. Since this technique is well suited for the functional characterization of electrogenic membrane transporters, it is expected to become a promising platform technology for drug screening and development.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Tadini-Buoninsegni F, Bartolommei G, Moncelli MR, Fendler K (2008) Charge transfer in P-type ATPases investigated on planar membranes. Arch Biochem Biophys 476:75–86
Ganea C, Fendler K (2009) Bacterial transporters: charge translocation and mechanism. Biochim Biophys Acta—Bioenerg 1787:706–713
Grewer C, Gameiro A, Mager T, Fendler K (2013) Electrophysiological characterization of membrane transport proteins. Annu Rev Biophys 42:95–120
Tadini-Buoninsegni F, Bartolommei G, Moncelli MR, Guidelli R, Inesi G (2006) Pre-steady state electrogenic events of Ca2+/H+ exchange and transport by the Ca2+-ATPase. J Biol Chem 281:37720–37727
Bartolommei G, Tadini-Buoninsegni F, Moncelli MR, Gemma S, Camodeca C, Butini S et al (2011) The Ca2+-ATPase (SERCA1) is inhibited by 4-aminoquinoline derivatives through interference with catalytic activation by Ca2+, whereas the ATPase E2 state remains functional. J Biol Chem 286:38383–38389
Kelety B, Diekert K, Tobien J, Watzke N, Dörner W, Obrdlik P et al (2006) Transporter assays using solid supported membranes: a novel screening platform for drug discovery. Assay Drug Dev Technol 4:575–582
Pintschovius J, Fendler K (1999) Charge translocation by the Na+/K+-ATPase investigated on solid supported membranes: rapid solution exchange with a new technique. Biophys J 76:814–826
Pintschovius J, Fendler K, Bamberg E (1999) Charge translocation by the Na+/K+-ATPase investigated on solid supported membranes: Cytoplasmic cation binding and release. Biophys J 76:827–836
Gramigni E, Tadini-Buoninsegni F, Bartolommei G, Santini G, Chelazzi G, Moncelli MR (2009) Inhibitory effect of Pb2+ on the transport cycle of the Na+, K+-ATPase. Chem Res Toxicol 22:1699–1704
Bartolommei G, Moncelli MR, Rispoli G, Kelety B, Tadini-Buoninsegni F (2009) Electrogenic ion pumps investigated on a solid supported membrane: comparison of current and voltage measurements. Langmuir 25:10925–10931
Tadini-Buoninsegni F, Pintschovius J, Cornelius F, Bamberg E, Fendler K (2000) K+ induced charge translocation in the phosphoenzyme formed from inorganic phosphate. In: Kaya S, Taniguchi K (eds) Na/K-ATPase and related ATPases. Elsevier, Amsterdam, pp 341–348
Tadini-Buoninsegni F, Bartolommei G, Moncelli MR, Pilankatta R, Lewis D, Inesi G (2010) ATP dependent charge movement in ATP7B Cu+-ATPase is demonstrated by pre-steady state electrical measurements. FEBS Lett 584:4619–4622
Tadini-Buoninsegni F, Bartolommei G, Moncelli MR, Inesi G, Galliani A, Sinisi M et al (2014) Translocation of platinum anticancer drugs by human copper ATPases ATP7A and ATP7B. Angew Chem Int Ed Engl 53:1297–1301
Seifert K, Fendler K, Bamberg E (1993) Charge transport by ion translocating membrane proteins on solid supported membranes. Biophys J 64:384–391
Schulz P, Garcia-Celma JJ, Fendler K (2008) SSM-based electrophysiology. Methods 46:97–103
Geibel S, Flores-Herr N, Licher T, Vollert H (2006) Establishment of cell-free electrophysiology for ion transporters: application for pharmacological profiling. J Biomol Screen 11:262–268
Garcia-Celma JJ, Hatahet L, Kunz W, Fendler K (2007) Specific anion and cation binding to lipid membranes investigated on a solid supported membrane. Langmuir 23:10074–10080
Bazzone A, Costa WS, Braner M, Călinescu O, Hatahet L, Fendler K (2013) Introduction to solid supported membrane based electrophysiology. J Vis Exp 75:e50230
Tadini Buoninsegni F, Bartolommei G, Moncelli MR, Inesi G, Guidelli R (2004) Time-resolved charge translocation by sarcoplasmic reticulum Ca-ATPase measured on a solid supported membrane. Biophys J 86:3671–3686
Acknowledgments
The authors acknowledge with gratitude financial support from the Italian Ministry of Education, University and Research (MIUR) and Ente Cassa di Risparmio di Firenze.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2016 Springer Science+Business Media New York
About this protocol
Cite this protocol
Tadini-Buoninsegni, F., Bartolommei, G. (2016). Electrophysiological Measurements on Solid Supported Membranes. In: Bublitz, M. (eds) P-Type ATPases. Methods in Molecular Biology, vol 1377. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3179-8_26
Download citation
DOI: https://doi.org/10.1007/978-1-4939-3179-8_26
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-3178-1
Online ISBN: 978-1-4939-3179-8
eBook Packages: Springer Protocols