TY - JOUR
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.
UR - http://www.scopus.com/inward/record.url?scp=85062634233&partnerID=8YFLogxK
U2 - 10.1042/BCJ20180891
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 -