Phosphatidylserine flipping by the P4-ATPase ATP8A2 is electrogenic

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Phosphatidylserine flipping by the P4-ATPase ATP8A2 is electrogenic. / Tadini-Buoninsegni, Francesco; Mikkelsen, Stine A; Mogensen, Louise S; Molday, Robert S.; Andersen, Jens Peter.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 116, No. 33, 13.08.2019, p. 16332-16337.

Research output: Contribution to journal/Conference contribution in journal/Contribution to newspaperJournal articleResearchpeer-review

Harvard

Tadini-Buoninsegni, F, Mikkelsen, SA, Mogensen, LS, Molday, RS & Andersen, JP 2019, 'Phosphatidylserine flipping by the P4-ATPase ATP8A2 is electrogenic', Proceedings of the National Academy of Sciences of the United States of America, vol. 116, no. 33, pp. 16332-16337. https://doi.org/10.1073/pnas.1910211116

APA

Tadini-Buoninsegni, F., Mikkelsen, S. A., Mogensen, L. S., Molday, R. S., & Andersen, J. P. (2019). Phosphatidylserine flipping by the P4-ATPase ATP8A2 is electrogenic. Proceedings of the National Academy of Sciences of the United States of America, 116(33), 16332-16337. https://doi.org/10.1073/pnas.1910211116

CBE

Tadini-Buoninsegni F, Mikkelsen SA, Mogensen LS, Molday RS, Andersen JP. 2019. Phosphatidylserine flipping by the P4-ATPase ATP8A2 is electrogenic. Proceedings of the National Academy of Sciences of the United States of America. 116(33):16332-16337. https://doi.org/10.1073/pnas.1910211116

MLA

Tadini-Buoninsegni, Francesco et al. "Phosphatidylserine flipping by the P4-ATPase ATP8A2 is electrogenic". Proceedings of the National Academy of Sciences of the United States of America. 2019, 116(33). 16332-16337. https://doi.org/10.1073/pnas.1910211116

Vancouver

Tadini-Buoninsegni F, Mikkelsen SA, Mogensen LS, Molday RS, Andersen JP. Phosphatidylserine flipping by the P4-ATPase ATP8A2 is electrogenic. Proceedings of the National Academy of Sciences of the United States of America. 2019 Aug 13;116(33):16332-16337. https://doi.org/10.1073/pnas.1910211116

Author

Tadini-Buoninsegni, Francesco ; Mikkelsen, Stine A ; Mogensen, Louise S ; Molday, Robert S. ; Andersen, Jens Peter. / Phosphatidylserine flipping by the P4-ATPase ATP8A2 is electrogenic. In: Proceedings of the National Academy of Sciences of the United States of America. 2019 ; Vol. 116, No. 33. pp. 16332-16337.

Bibtex

@article{ad73d2f357874ac1bbcb777134f3f740,
title = "Phosphatidylserine flipping by the P4-ATPase ATP8A2 is electrogenic",
abstract = "Phospholipid flippases (P4-ATPases) utilize ATP to translocate specific phospholipids from the exoplasmic leaflet to the cytoplasmic leaflet of biological membranes, thus generating and maintaining transmembrane lipid asymmetry essential for a variety of cellular processes. P4-ATPases belong to the P-type ATPase protein family, which also encompasses the ion transporting P2-ATPases: Ca2+-ATPase, Na+,K+-ATPase, and H+,K+-ATPase. In comparison with the P2-ATPases, understanding of P4-ATPases is still very limited. The electrogenicity of P4-ATPases has not been explored, and it is not known whether lipid transfer between membrane bilayer leaflets can lead to displacement of charge across the membrane. A related question is whether P4-ATPases countertransport ions or other substrates in the opposite direction, similar to the P2-ATPases. Using an electrophysiological method based on solid supported membranes, we observed the generation of a transient electrical current by the mammalian P4-ATPase ATP8A2 in the presence of ATP and the negatively charged lipid substrate phosphatidylserine, whereas only a diminutive current was generated with the lipid substrate phosphatidylethanolamine, which carries no or little charge under the conditions of the measurement. The current transient seen with phosphatidylserine was abolished by the mutation E198Q, which blocks dephosphorylation. Likewise, mutation I364M, which causes the neurological disorder cerebellar ataxia, mental retardation, and disequilibrium (CAMRQ) syndrome, strongly interfered with the electrogenic lipid translocation. It is concluded that the electrogenicity is associated with a step in the ATPase reaction cycle directly involved in translocation of the lipid. These measurements also showed that no charged substrate is being countertransported, thereby distinguishing the P4-ATPase from P2-ATPases.",
author = "Francesco Tadini-Buoninsegni and Mikkelsen, {Stine A} and Mogensen, {Louise S} and Molday, {Robert S.} and Andersen, {Jens Peter}",
year = "2019",
month = "8",
day = "13",
doi = "10.1073/pnas.1910211116",
language = "English",
volume = "116",
pages = "16332--16337",
journal = "Proceedings of the National Academy of Sciences of the United States of America",
issn = "0027-8424",
publisher = "The National Academy of Sciences of the United States of America",
number = "33",

}

RIS

TY - JOUR

T1 - Phosphatidylserine flipping by the P4-ATPase ATP8A2 is electrogenic

AU - Tadini-Buoninsegni, Francesco

AU - Mikkelsen, Stine A

AU - Mogensen, Louise S

AU - Molday, Robert S.

AU - Andersen, Jens Peter

PY - 2019/8/13

Y1 - 2019/8/13

N2 - Phospholipid flippases (P4-ATPases) utilize ATP to translocate specific phospholipids from the exoplasmic leaflet to the cytoplasmic leaflet of biological membranes, thus generating and maintaining transmembrane lipid asymmetry essential for a variety of cellular processes. P4-ATPases belong to the P-type ATPase protein family, which also encompasses the ion transporting P2-ATPases: Ca2+-ATPase, Na+,K+-ATPase, and H+,K+-ATPase. In comparison with the P2-ATPases, understanding of P4-ATPases is still very limited. The electrogenicity of P4-ATPases has not been explored, and it is not known whether lipid transfer between membrane bilayer leaflets can lead to displacement of charge across the membrane. A related question is whether P4-ATPases countertransport ions or other substrates in the opposite direction, similar to the P2-ATPases. Using an electrophysiological method based on solid supported membranes, we observed the generation of a transient electrical current by the mammalian P4-ATPase ATP8A2 in the presence of ATP and the negatively charged lipid substrate phosphatidylserine, whereas only a diminutive current was generated with the lipid substrate phosphatidylethanolamine, which carries no or little charge under the conditions of the measurement. The current transient seen with phosphatidylserine was abolished by the mutation E198Q, which blocks dephosphorylation. Likewise, mutation I364M, which causes the neurological disorder cerebellar ataxia, mental retardation, and disequilibrium (CAMRQ) syndrome, strongly interfered with the electrogenic lipid translocation. It is concluded that the electrogenicity is associated with a step in the ATPase reaction cycle directly involved in translocation of the lipid. These measurements also showed that no charged substrate is being countertransported, thereby distinguishing the P4-ATPase from P2-ATPases.

AB - Phospholipid flippases (P4-ATPases) utilize ATP to translocate specific phospholipids from the exoplasmic leaflet to the cytoplasmic leaflet of biological membranes, thus generating and maintaining transmembrane lipid asymmetry essential for a variety of cellular processes. P4-ATPases belong to the P-type ATPase protein family, which also encompasses the ion transporting P2-ATPases: Ca2+-ATPase, Na+,K+-ATPase, and H+,K+-ATPase. In comparison with the P2-ATPases, understanding of P4-ATPases is still very limited. The electrogenicity of P4-ATPases has not been explored, and it is not known whether lipid transfer between membrane bilayer leaflets can lead to displacement of charge across the membrane. A related question is whether P4-ATPases countertransport ions or other substrates in the opposite direction, similar to the P2-ATPases. Using an electrophysiological method based on solid supported membranes, we observed the generation of a transient electrical current by the mammalian P4-ATPase ATP8A2 in the presence of ATP and the negatively charged lipid substrate phosphatidylserine, whereas only a diminutive current was generated with the lipid substrate phosphatidylethanolamine, which carries no or little charge under the conditions of the measurement. The current transient seen with phosphatidylserine was abolished by the mutation E198Q, which blocks dephosphorylation. Likewise, mutation I364M, which causes the neurological disorder cerebellar ataxia, mental retardation, and disequilibrium (CAMRQ) syndrome, strongly interfered with the electrogenic lipid translocation. It is concluded that the electrogenicity is associated with a step in the ATPase reaction cycle directly involved in translocation of the lipid. These measurements also showed that no charged substrate is being countertransported, thereby distinguishing the P4-ATPase from P2-ATPases.

U2 - 10.1073/pnas.1910211116

DO - 10.1073/pnas.1910211116

M3 - Journal article

C2 - 31371510

VL - 116

SP - 16332

EP - 16337

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

SN - 0027-8424

IS - 33

ER -