The lipid head group is the key element for substrate recognition by the P4 ATPase ALA2: A phosphatidylserine flippase

Lisa Theorin; Kristina Faxén; Danny Mollerup Sørensen; Rebekka Migotti; Gunnar Dittmar; Jürgen Schiller; David L. Daleke; Michael Palmgren; Rosa Laura López-Marqués; Thomas Günther Pomorski

doi:10.1042/BCJ20180891

The lipid head group is the key element for substrate recognition by the P4 ATPase ALA2: A phosphatidylserine flippase

Lisa Theorin, Kristina Faxén, Danny Mollerup Sørensen, Rebekka Migotti, Gunnar Dittmar, Jürgen Schiller, David L. Daleke, Michael Palmgren, Rosa Laura López-Marqués^*, Thomas Günther Pomorski

^*Corresponding author for this work

Section for Transport Biology

2 Citations (Scopus)

22 Downloads (Pure)

Abstract

Type IV P-type ATPases (P4 ATPases) are lipid flippases that catalyze phospholipid transport from the exoplasmic to the cytoplasmic leaflet of cellular membranes, but the mechanism by which they recognize and transport phospholipids through the lipid bilayer remains unknown. In the present study, we succeeded in purifying recombinant aminophospholipid ATPase 2 (ALA2), a member of the P4 ATPase subfamily in Arabidopsis thaliana, in complex with the ALA-interacting subunit 5 (ALIS5). The ATP hydrolytic activity of the ALA2–ALIS5 complex was stimulated in a highly specific manner by phosphatidylserine. Small changes in the stereochemistry or the functional groups of the phosphatidylserine head group affected enzymatic activity, whereas alteration in the length and composition of the acyl chains only had minor effects. Likewise, the enzymatic activity of the ALA2–ALIS5 complex was stimulated by both mono- and di-acyl phospha-tidylserines. Taken together, the results identify the lipid head group as the key structural element for substrate recognition by the P4 ATPase.

Original language	English
Journal	Biochemical Journal
Volume	476
Issue number	5
Pages (from-to)	783-794
ISSN	0264-6021
DOIs	https://doi.org/10.1042/BCJ20180891
Publication status	Published - 6 Mar 2019

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The lipid head group is the key element forsubstrate recognition by the P4 ATPaseALA2Final published version, 8.56 MBLicence: CC BY-NC-ND

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title = "The lipid head group is the key element for substrate recognition by the P4 ATPase ALA2: A phosphatidylserine flippase",

abstract = "Type IV P-type ATPases (P4 ATPases) are lipid flippases that catalyze phospholipid transport from the exoplasmic to the cytoplasmic leaflet of cellular membranes, but the mechanism by which they recognize and transport phospholipids through the lipid bilayer remains unknown. In the present study, we succeeded in purifying recombinant aminophospholipid ATPase 2 (ALA2), a member of the P4 ATPase subfamily in Arabidopsis thaliana, in complex with the ALA-interacting subunit 5 (ALIS5). The ATP hydrolytic activity of the ALA2–ALIS5 complex was stimulated in a highly specific manner by phosphatidylserine. Small changes in the stereochemistry or the functional groups of the phosphatidylserine head group affected enzymatic activity, whereas alteration in the length and composition of the acyl chains only had minor effects. Likewise, the enzymatic activity of the ALA2–ALIS5 complex was stimulated by both mono- and di-acyl phospha-tidylserines. Taken together, the results identify the lipid head group as the key structural element for substrate recognition by the P4 ATPase.",

author = "Lisa Theorin and Kristina Fax{\'e}n and S{\o}rensen, {Danny Mollerup} and Rebekka Migotti and Gunnar Dittmar and J{\"u}rgen Schiller and Daleke, {David L.} and Michael Palmgren and L{\'o}pez-Marqu{\'e}s, {Rosa Laura} and Pomorski, {Thomas G{\"u}nther}",

year = "2019",

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day = "6",

doi = "10.1042/BCJ20180891",

language = "English",

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pages = "783--794",

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T1 - The lipid head group is the key element for substrate recognition by the P4 ATPase ALA2

T2 - A phosphatidylserine flippase

AU - Theorin, Lisa

AU - Faxén, Kristina

AU - Sørensen, Danny Mollerup

AU - Migotti, Rebekka

AU - Dittmar, Gunnar

AU - Schiller, Jürgen

AU - Daleke, David L.

AU - Palmgren, Michael

AU - López-Marqués, Rosa Laura

AU - Pomorski, Thomas Günther

PY - 2019/3/6

Y1 - 2019/3/6

N2 - Type IV P-type ATPases (P4 ATPases) are lipid flippases that catalyze phospholipid transport from the exoplasmic to the cytoplasmic leaflet of cellular membranes, but the mechanism by which they recognize and transport phospholipids through the lipid bilayer remains unknown. In the present study, we succeeded in purifying recombinant aminophospholipid ATPase 2 (ALA2), a member of the P4 ATPase subfamily in Arabidopsis thaliana, in complex with the ALA-interacting subunit 5 (ALIS5). The ATP hydrolytic activity of the ALA2–ALIS5 complex was stimulated in a highly specific manner by phosphatidylserine. Small changes in the stereochemistry or the functional groups of the phosphatidylserine head group affected enzymatic activity, whereas alteration in the length and composition of the acyl chains only had minor effects. Likewise, the enzymatic activity of the ALA2–ALIS5 complex was stimulated by both mono- and di-acyl phospha-tidylserines. Taken together, the results identify the lipid head group as the key structural element for substrate recognition by the P4 ATPase.

AB - Type IV P-type ATPases (P4 ATPases) are lipid flippases that catalyze phospholipid transport from the exoplasmic to the cytoplasmic leaflet of cellular membranes, but the mechanism by which they recognize and transport phospholipids through the lipid bilayer remains unknown. In the present study, we succeeded in purifying recombinant aminophospholipid ATPase 2 (ALA2), a member of the P4 ATPase subfamily in Arabidopsis thaliana, in complex with the ALA-interacting subunit 5 (ALIS5). The ATP hydrolytic activity of the ALA2–ALIS5 complex was stimulated in a highly specific manner by phosphatidylserine. Small changes in the stereochemistry or the functional groups of the phosphatidylserine head group affected enzymatic activity, whereas alteration in the length and composition of the acyl chains only had minor effects. Likewise, the enzymatic activity of the ALA2–ALIS5 complex was stimulated by both mono- and di-acyl phospha-tidylserines. Taken together, the results identify the lipid head group as the key structural element for substrate recognition by the P4 ATPase.

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DO - 10.1042/BCJ20180891

M3 - Journal article

C2 - 30755463

AN - SCOPUS:85062634233

SN - 0264-6021

VL - 476

SP - 783

EP - 794

JO - Biochemical Journal

JF - Biochemical Journal

IS - 5

ER -

The lipid head group is the key element for substrate recognition by the P4 ATPase ALA2: A phosphatidylserine flippase

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