Role of post-translational modifications at the β-subunit ectodomain in complex association with a promiscuous plant P4-ATPase
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Role of post-translational modifications at the β-subunit ectodomain in complex association with a promiscuous plant P4-ATPase. / Costa, Sara; Marek, Magdalena; Axelsen, Kristian Buhl; Theorin, Lisa; Günther-Pomorski, Thomas; Lopez Marques, Rosa Laura.
In: Biochemical Journal, Vol. 473, No. 11, 2016, p. 1605-1615.Research output: Contribution to journal › Journal article › Research › peer-review
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T1 - Role of post-translational modifications at the β-subunit ectodomain in complex association with a promiscuous plant P4-ATPase
AU - Costa, Sara
AU - Marek, Magdalena
AU - Axelsen, Kristian Buhl
AU - Theorin, Lisa
AU - Günther-Pomorski, Thomas
AU - Lopez Marques, Rosa Laura
N1 - © 2016 The Author(s). published by Portland Press Limited on behalf of the Biochemical Society.
PY - 2016
Y1 - 2016
N2 - P-type ATPases of subfamily IV (P4-ATPases) constitute a major group of phospholipid flippases that form heteromeric complexes with members of the Cdc50 (cell division control 50) protein family. Some P4-ATPases interact specifically with only one β-subunit isoform, whereas others are promiscuous and can interact with several isoforms. In the present study, we used a site-directed mutagenesis approach to assess the role of post-translational modifications at the plant ALIS5 β-subunit ectodomain in the functionality of the promiscuous plant P4-ATPase ALA2. We identified two N-glycosylated residues, Asn(181) and Asn(231) Whereas mutation of Asn(231) seems to have a small effect on P4-ATPase complex formation, mutation of evolutionarily conserved Asn(181) disrupts interaction between the two subunits. Of the four cysteine residues located in the ALIS5 ectodomain, mutation of Cys(86) and Cys(107) compromises complex association, but the mutant β-subunits still promote complex trafficking and activity to some extent. In contrast, disruption of a conserved disulfide bond between Cys(158) and Cys(172) has no effect on the P4-ATPase complex. Our results demonstrate that post-translational modifications in the β-subunit have different functional roles in different organisms, which may be related to the promiscuity of the P4-ATPase.
AB - P-type ATPases of subfamily IV (P4-ATPases) constitute a major group of phospholipid flippases that form heteromeric complexes with members of the Cdc50 (cell division control 50) protein family. Some P4-ATPases interact specifically with only one β-subunit isoform, whereas others are promiscuous and can interact with several isoforms. In the present study, we used a site-directed mutagenesis approach to assess the role of post-translational modifications at the plant ALIS5 β-subunit ectodomain in the functionality of the promiscuous plant P4-ATPase ALA2. We identified two N-glycosylated residues, Asn(181) and Asn(231) Whereas mutation of Asn(231) seems to have a small effect on P4-ATPase complex formation, mutation of evolutionarily conserved Asn(181) disrupts interaction between the two subunits. Of the four cysteine residues located in the ALIS5 ectodomain, mutation of Cys(86) and Cys(107) compromises complex association, but the mutant β-subunits still promote complex trafficking and activity to some extent. In contrast, disruption of a conserved disulfide bond between Cys(158) and Cys(172) has no effect on the P4-ATPase complex. Our results demonstrate that post-translational modifications in the β-subunit have different functional roles in different organisms, which may be related to the promiscuity of the P4-ATPase.
KW - Journal Article
U2 - 10.1042/BCJ20160207
DO - 10.1042/BCJ20160207
M3 - Journal article
C2 - 27048590
VL - 473
SP - 1605
EP - 1615
JO - Biochemical Journal
JF - Biochemical Journal
SN - 0264-6021
IS - 11
ER -
ID: 169135801