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Type IIA calcium ATPases of arabidopsis thaliana-their organ and membrane distributions

Type IIA calcium ATPases of arabidopsis thaliana-their organ and membrane distributions
Type IIA calcium ATPases of arabidopsis thaliana-their organ and membrane distributions

The membrane and tissue distributions of Type IIA calcium ATPases in highers plants were investigated using specific anti-peptide antibodies against two Arabidopsis thaliana Type IIA calcium ATPases, ACA3 (AtECA1) and ACA5 (AtECA2). The antibodies were raised against a region of the predicted M7-M8 lumenal loop of these proteins. Specific antibodies against ACA3 that had been produced prior to the start of this study were characterised, and, as part of this study, antibodies were raised against ACA5. In addition, several antibodies against different regions of a mammalian Type IIA calcium ATPase, the rabbit SERCA 1, were used to investigate the occurrence of a third Arabidopsis Type IIA calcium ATPase, ACA6 (AtECA3), and to investigate whether further Type IIA calcium ATPases might occur in Arabidopsis.

ACA3 has a predicted molecular weight of 116.4 kD, and the antibody against ACA3 detected a protein of 116 kD on western blots of membrane proteins separated by SDS-PAGE. ACA5 has a predicted molecular weight of 115.8 kD, and the antibody against ACA5 detected protein at molecular weights of 115 and 111 kD. Most of the antibodies raised against the SERCA 1 that were predicted to react specifically with Type IIA calcium ATPases of higher plants detected protein at around 116 kD.

Membranes were isolated from Arabidopsis thaliana roots, leaf, stem, flower and silique tissues, and immunoblotting indicated that ACA3 and ACA5 were particularly abundant in stem membranes, with lower levels in silique, then leaf, and then flower membranes. Lower levels of ACA3 were detected in root membranes than in membranes from aerial organs, whilst ACA5 was not detected in root membranes. An anti-peptide antibody against the SERCA 1 that is likely to detect ACA6 (predicted molecular weight 109.0 kD) but not ACA3 or ACA5, and may also detect other Type IIA ATPases, detected protein of 116 kD and also of other sizes.

University of Southampton
Mills, Rebecca Fay
a0a792e8-e2b3-468c-aa5c-2e273fd29f56
Mills, Rebecca Fay
a0a792e8-e2b3-468c-aa5c-2e273fd29f56

Mills, Rebecca Fay (1999) Type IIA calcium ATPases of arabidopsis thaliana-their organ and membrane distributions. University of Southampton, Doctoral Thesis.

Record type: Thesis (Doctoral)

Abstract

The membrane and tissue distributions of Type IIA calcium ATPases in highers plants were investigated using specific anti-peptide antibodies against two Arabidopsis thaliana Type IIA calcium ATPases, ACA3 (AtECA1) and ACA5 (AtECA2). The antibodies were raised against a region of the predicted M7-M8 lumenal loop of these proteins. Specific antibodies against ACA3 that had been produced prior to the start of this study were characterised, and, as part of this study, antibodies were raised against ACA5. In addition, several antibodies against different regions of a mammalian Type IIA calcium ATPase, the rabbit SERCA 1, were used to investigate the occurrence of a third Arabidopsis Type IIA calcium ATPase, ACA6 (AtECA3), and to investigate whether further Type IIA calcium ATPases might occur in Arabidopsis.

ACA3 has a predicted molecular weight of 116.4 kD, and the antibody against ACA3 detected a protein of 116 kD on western blots of membrane proteins separated by SDS-PAGE. ACA5 has a predicted molecular weight of 115.8 kD, and the antibody against ACA5 detected protein at molecular weights of 115 and 111 kD. Most of the antibodies raised against the SERCA 1 that were predicted to react specifically with Type IIA calcium ATPases of higher plants detected protein at around 116 kD.

Membranes were isolated from Arabidopsis thaliana roots, leaf, stem, flower and silique tissues, and immunoblotting indicated that ACA3 and ACA5 were particularly abundant in stem membranes, with lower levels in silique, then leaf, and then flower membranes. Lower levels of ACA3 were detected in root membranes than in membranes from aerial organs, whilst ACA5 was not detected in root membranes. An anti-peptide antibody against the SERCA 1 that is likely to detect ACA6 (predicted molecular weight 109.0 kD) but not ACA3 or ACA5, and may also detect other Type IIA ATPases, detected protein of 116 kD and also of other sizes.

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Published date: 1999

Identifiers

Local EPrints ID: 464003
URI: http://eprints.soton.ac.uk/id/eprint/464003
PURE UUID: a231d0aa-5721-4a13-95b6-ac9c357d4c2d

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Date deposited: 04 Jul 2022 21:00
Last modified: 04 Jul 2022 21:00

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Contributors

Author: Rebecca Fay Mills

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