Interaction of N-terminal peptide analogues of the Na+,K+-ATPase with membranes

Nguyen, K; Garcia, A; Sani, MA; Diaz, D; Dubey, V; Clayton, D; Dal Poggetto, G; Cornelius, F; Payne, RJ; Separovic, F; Khandelia, H; Clarke, RJ

Interaction of N-terminal peptide analogues of the Na+,K+-ATPase with membranes

Nguyen, K Garcia, A

Sani, MA Diaz, D Dubey, V Clayton, D Dal Poggetto, G Cornelius, F Payne, RJ Separovic, F Khandelia, H Clarke, RJ

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Publication Type:: Journal Article
Citation:: Biochimica et Biophysica Acta - Biomembranes, 2018, 1860 (6), pp. 1282 - 1291
Issue Date:: 2018-06-01

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Field	Value	Language
dc.contributor.author	Nguyen, K	en_US
dc.contributor.author	Garcia, A https://orcid.org/0000-0002-1159-4567	en_US
dc.contributor.author	Sani, MA	en_US
dc.contributor.author	Diaz, D	en_US
dc.contributor.author	Dubey, V	en_US
dc.contributor.author	Clayton, D	en_US
dc.contributor.author	Dal Poggetto, G	en_US
dc.contributor.author	Cornelius, F	en_US
dc.contributor.author	Payne, RJ	en_US
dc.contributor.author	Separovic, F	en_US
dc.contributor.author	Khandelia, H	en_US
dc.contributor.author	Clarke, RJ	en_US
dc.date.available	2018-03-05	en_US
dc.date.issued	2018-06-01	en_US
dc.identifier.citation	Biochimica et Biophysica Acta - Biomembranes, 2018, 1860 (6), pp. 1282 - 1291	en_US
dc.identifier.issn	0005-2736	en_US
dc.identifier.uri	http://hdl.handle.net/10453/134041
dc.description.abstract	© 2018 The Na+,K+-ATPase, which is present in the plasma membrane of all animal cells, plays a crucial role in maintaining the Na+ and K+ electrochemical potential gradients across the membrane. Recent studies have suggested that the N-terminus of the protein's catalytic α-subunit is involved in an electrostatic interaction with the surrounding membrane, which controls the protein's conformational equilibrium. However, because the N-terminus could not yet be resolved in any X-ray crystal structures, little information about this interaction is so far available. In measurements utilising poly-L-lysine as a model of the protein's lysine-rich N-terminus and using lipid vesicles of defined composition, here we have identified the most likely origin of the interaction as one between positively charged lysine residues of the N-terminus and negatively charged headgroups of phospholipids (notably phosphatidylserine) in the surrounding membrane. Furthermore, to isolate which segments of the N-terminus could be involved in membrane binding, we chemically synthesized N-terminal fragments of various lengths. Based on a combination of results from RH421 UV/visible absorbance measurements and solid-state 31P and 2H NMR using these N-terminal fragments as well as MD simulations it appears that the membrane interaction arises from lysine residues prior to the conserved LKKE motif of the N-terminus. The MD simulations indicate that the strength of the interaction varies significantly between different enzyme conformations.	en_US
dc.relation.ispartof	Biochimica et Biophysica Acta - Biomembranes	en_US
dc.relation.isbasedon	10.1016/j.bbamem.2018.03.002	en_US
dc.subject.classification	Biophysics	en_US
dc.subject.classification	Biochemistry & Molecular Biology	en_US
dc.subject.mesh	Cell Membrane	en_US
dc.subject.mesh	Animals	en_US
dc.subject.mesh	Sus scrofa	en_US
dc.subject.mesh	Polylysine	en_US
dc.subject.mesh	Peptide Fragments	en_US
dc.subject.mesh	Spectrophotometry, Ultraviolet	en_US
dc.subject.mesh	Nuclear Magnetic Resonance, Biomolecular	en_US
dc.subject.mesh	Sequence Alignment	en_US
dc.subject.mesh	Amino Acid Sequence	en_US
dc.subject.mesh	Conserved Sequence	en_US
dc.subject.mesh	Protein Conformation	en_US
dc.subject.mesh	Sequence Homology, Amino Acid	en_US
dc.subject.mesh	Sodium-Potassium-Exchanging ATPase	en_US
dc.subject.mesh	Molecular Docking Simulation	en_US
dc.title	Interaction of N-terminal peptide analogues of the Na<sup>+</sup>,K<sup>+</sup>-ATPase with membranes	en_US
dc.type	Journal Article
utslib.citation.volume	6	en_US
utslib.citation.volume	1860	en_US
utslib.for	0601 Biochemistry and Cell Biology	en_US
utslib.for	0699 Other Biological Sciences	en_US
utslib.for	0904 Chemical Engineering	en_US
pubs.embargo.period	Not known	en_US
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
pubs.organisational-group	/University of Technology Sydney/Faculty of Science/School of Life Sciences
utslib.copyright.status	closed_access
pubs.issue	6	en_US
pubs.publication-status	Published	en_US
pubs.volume	1860	en_US

Abstract:

© 2018 The Na+,K+-ATPase, which is present in the plasma membrane of all animal cells, plays a crucial role in maintaining the Na+ and K+ electrochemical potential gradients across the membrane. Recent studies have suggested that the N-terminus of the protein's catalytic α-subunit is involved in an electrostatic interaction with the surrounding membrane, which controls the protein's conformational equilibrium. However, because the N-terminus could not yet be resolved in any X-ray crystal structures, little information about this interaction is so far available. In measurements utilising poly-L-lysine as a model of the protein's lysine-rich N-terminus and using lipid vesicles of defined composition, here we have identified the most likely origin of the interaction as one between positively charged lysine residues of the N-terminus and negatively charged headgroups of phospholipids (notably phosphatidylserine) in the surrounding membrane. Furthermore, to isolate which segments of the N-terminus could be involved in membrane binding, we chemically synthesized N-terminal fragments of various lengths. Based on a combination of results from RH421 UV/visible absorbance measurements and solid-state 31P and 2H NMR using these N-terminal fragments as well as MD simulations it appears that the membrane interaction arises from lysine residues prior to the conserved LKKE motif of the N-terminus. The MD simulations indicate that the strength of the interaction varies significantly between different enzyme conformations.

Please use this identifier to cite or link to this item:

http://hdl.handle.net/10453/134041

Interaction of N-terminal peptide analogues of the Na<sup>+</sup>,K<sup>+</sup>-ATPase with membranes