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Biochem J. 2019 Mar 6;476(5):783-794. doi: 10.1042/BCJ20180891.

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

Author information

1
Department of Plant and Environmental Sciences, University of Copenhagen, Copenhagen, Denmark.
2
Max-Delbrück Centrum for Molecular Medicine, Berlin, Germany.
3
Proteome and Genome Research Laboratory, Luxembourg Institute of Health, Strassen, Luxembourg.
4
Institute of Medical Physics and Biophysics, University of Leipzig, Leipzig, Germany.
5
Medical Sciences/Department of Biochemistry and Molecular Biology, Indiana University School of Medicine, Bloomington, IN, U.S.A.
6
Department of Plant and Environmental Sciences, University of Copenhagen, Copenhagen, Denmark rlo@plen.ku.dk Thomas.Guenther-Pomorski@rub.de.
7
Department of Molecular Biochemistry, Faculty of Chemistry and Biochemistry, Ruhr University Bochum, Bochum, Germany.

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 phosphatidylserines. Taken together, the results identify the lipid head group as the key structural element for substrate recognition by the P4 ATPase.

KEYWORDS:

Arabidopsis thaliana; FLAG-tag purification; lipid flippase; lysophospholipid; membrane protein complex

PMID:
30755463
PMCID:
PMC6402034
DOI:
10.1042/BCJ20180891
[Indexed for MEDLINE]
Free PMC Article

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