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Characterization of a Listeria monocytogenes Ca(2+) pump : a SERCA-type ATPase with only one Ca(2+)-binding site

Faxén, Kristina; Andersen, Jacob Lauwring; Gourdon, Pontus LU ; Fedosova, Natalya; Morth, Jens Preben; Nissen, Poul and Møller, Jesper Vuust (2011) In Journal of Biological Chemistry 286(2). p.17-1609
Abstract

We have characterized a putative Ca(2+)-ATPase from the pathogenic bacterium Listeria monocytogenes with the locus tag lmo0841. The purified and detergent-solubilized protein, which we have named Listeria monocytogenes Ca(2+)-ATPase 1 (LMCA1), performs a Ca(2+)-dependent ATP hydrolysis and actively transports Ca(2+) after reconstitution in dioleoylphosphatidyl-choline vesicles. Despite a high sequence similarity to the sarcoplasmic reticulum Ca(2+)-ATPase (SERCA1a) and plasma membrane Ca(2+)-ATPase (PMCA), LMCA1 exhibits important biochemical differences such as a low Ca(2+) affinity (K(0.5) ∼80 μm) and a high pH optimum (pH ∼9). Mutational studies indicate that the unusually high pH optimum can be partially ascribed to the presence of... (More)

We have characterized a putative Ca(2+)-ATPase from the pathogenic bacterium Listeria monocytogenes with the locus tag lmo0841. The purified and detergent-solubilized protein, which we have named Listeria monocytogenes Ca(2+)-ATPase 1 (LMCA1), performs a Ca(2+)-dependent ATP hydrolysis and actively transports Ca(2+) after reconstitution in dioleoylphosphatidyl-choline vesicles. Despite a high sequence similarity to the sarcoplasmic reticulum Ca(2+)-ATPase (SERCA1a) and plasma membrane Ca(2+)-ATPase (PMCA), LMCA1 exhibits important biochemical differences such as a low Ca(2+) affinity (K(0.5) ∼80 μm) and a high pH optimum (pH ∼9). Mutational studies indicate that the unusually high pH optimum can be partially ascribed to the presence of an arginine residue (Arg-795), corresponding in sequence alignments to the Glu-908 position at Ca(2+) binding site I of rabbit SERCA1a, but probably with an exposed position in LMCA1. The arginine is characteristic of a large group of putative bacterial Ca(2+)-ATPases. Moreover, we demonstrate that H(+) is countertransported with a transport stoichiometry of 1 Ca(2+) out and 1 H(+) in per ATP hydrolyzed. The ATPase may serve an important function by removing Ca(2+) from the microorganism in environmental conditions when e.g. stressed by high Ca(2+) and alkaline pH.

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publishing date
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published
keywords
Adenosine Triphosphate, Alkalies, Amino Acid Sequence, Animals, Binding Sites, Biological Transport, Active, Calcium, Electrochemical Techniques, Hydrogen-Ion Concentration, Listeria monocytogenes, Molecular Sequence Data, Muscle, Skeletal, Protein Structure, Tertiary, Rabbits, Sarcoplasmic Reticulum Calcium-Transporting ATPases, Journal Article, Research Support, Non-U.S. Gov't
in
Journal of Biological Chemistry
volume
286
issue
2
pages
17 - 1609
publisher
ASBMB
external identifiers
  • scopus:78651394444
ISSN
1083-351X
DOI
10.1074/jbc.M110.176784
language
English
LU publication?
no
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4a6564be-279d-4d66-8502-b967dba89ebf
date added to LUP
2017-04-29 15:31:54
date last changed
2017-05-07 04:52:51
@article{4a6564be-279d-4d66-8502-b967dba89ebf,
  abstract     = {<p>We have characterized a putative Ca(2+)-ATPase from the pathogenic bacterium Listeria monocytogenes with the locus tag lmo0841. The purified and detergent-solubilized protein, which we have named Listeria monocytogenes Ca(2+)-ATPase 1 (LMCA1), performs a Ca(2+)-dependent ATP hydrolysis and actively transports Ca(2+) after reconstitution in dioleoylphosphatidyl-choline vesicles. Despite a high sequence similarity to the sarcoplasmic reticulum Ca(2+)-ATPase (SERCA1a) and plasma membrane Ca(2+)-ATPase (PMCA), LMCA1 exhibits important biochemical differences such as a low Ca(2+) affinity (K(0.5) ∼80 μm) and a high pH optimum (pH ∼9). Mutational studies indicate that the unusually high pH optimum can be partially ascribed to the presence of an arginine residue (Arg-795), corresponding in sequence alignments to the Glu-908 position at Ca(2+) binding site I of rabbit SERCA1a, but probably with an exposed position in LMCA1. The arginine is characteristic of a large group of putative bacterial Ca(2+)-ATPases. Moreover, we demonstrate that H(+) is countertransported with a transport stoichiometry of 1 Ca(2+) out and 1 H(+) in per ATP hydrolyzed. The ATPase may serve an important function by removing Ca(2+) from the microorganism in environmental conditions when e.g. stressed by high Ca(2+) and alkaline pH.</p>},
  author       = {Faxén, Kristina and Andersen, Jacob Lauwring and Gourdon, Pontus and Fedosova, Natalya and Morth, Jens Preben and Nissen, Poul and Møller, Jesper Vuust},
  issn         = {1083-351X},
  keyword      = {Adenosine Triphosphate,Alkalies,Amino Acid Sequence,Animals,Binding Sites,Biological Transport, Active,Calcium,Electrochemical Techniques,Hydrogen-Ion Concentration,Listeria monocytogenes,Molecular Sequence Data,Muscle, Skeletal,Protein Structure, Tertiary,Rabbits,Sarcoplasmic Reticulum Calcium-Transporting ATPases,Journal Article,Research Support, Non-U.S. Gov't},
  language     = {eng},
  month        = {01},
  number       = {2},
  pages        = {17--1609},
  publisher    = {ASBMB},
  series       = {Journal of Biological Chemistry},
  title        = {Characterization of a Listeria monocytogenes Ca(2+) pump : a SERCA-type ATPase with only one Ca(2+)-binding site},
  url          = {http://dx.doi.org/10.1074/jbc.M110.176784},
  volume       = {286},
  year         = {2011},
}