Characterization of PISP, a novel single-PDZ protein that binds to all plasma membrane Ca2+-ATPase b-splice variants

Geoffrey M. Goellner, Steven J. DeMarco, Emanuel E. Strehler

Research output: Contribution to journalArticle

49 Citations (Scopus)

Abstract

Plasma membrane Ca2+ ATPases (PMCAs) maintain intracellular Ca2+ homeostasis and participate in the local regulation of Ca2+ signaling. Spatially separate demands for Ca2+ regulation require proper membrane targeting of PMCAs, but the mechanism of PMCA targeting is unknown. Using the PMCA2b carboxyl-terminal tail as yeast two-hybrid bait, we isolated a novel PDZ domain-containing protein from a human brain cDNA library. This protein, named PISP for PMCA-interacting single-PDZ protein, consists of 140 amino acids and contains little else besides a single PDZ domain. Pulldown experiments showed that PISP interacts with all PMCA b-splice forms. PISP was found to be ubiquitously expressed and, in MDCK cells, was present in a punctate pattern throughout the cytosol and at the basolateral membrane. When added to microsomal membranes expressing PMCA4b, PISP was unable to stimulate the PMCA-dependent ATPase activity. Our data suggest that PISP is a transiently interacting partner of the PMCA b-splice forms that may play a role in their sorting to or from the plasma membrane.

Original languageEnglish (US)
Pages (from-to)461-471
Number of pages11
JournalAnnals of the New York Academy of Sciences
Volume986
StatePublished - 2003

Fingerprint

Calcium-Transporting ATPases
Cell membranes
Cell Membrane
Proteins
PDZ Domains
Membranes
Madin Darby Canine Kidney Cells
Membrane
Protein
Plasma
Gene Library
Sorting
Cytosol
Yeast
Adenosine Triphosphatases
Tail
Brain
Homeostasis
Yeasts
Amino Acids

Keywords

  • Calcium pump
  • PDZ domain
  • Plasma membrane Ca-ATPase
  • PMCA

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Characterization of PISP, a novel single-PDZ protein that binds to all plasma membrane Ca2+-ATPase b-splice variants. / Goellner, Geoffrey M.; DeMarco, Steven J.; Strehler, Emanuel E.

In: Annals of the New York Academy of Sciences, Vol. 986, 2003, p. 461-471.

Research output: Contribution to journalArticle

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AU - Strehler, Emanuel E.

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AB - Plasma membrane Ca2+ ATPases (PMCAs) maintain intracellular Ca2+ homeostasis and participate in the local regulation of Ca2+ signaling. Spatially separate demands for Ca2+ regulation require proper membrane targeting of PMCAs, but the mechanism of PMCA targeting is unknown. Using the PMCA2b carboxyl-terminal tail as yeast two-hybrid bait, we isolated a novel PDZ domain-containing protein from a human brain cDNA library. This protein, named PISP for PMCA-interacting single-PDZ protein, consists of 140 amino acids and contains little else besides a single PDZ domain. Pulldown experiments showed that PISP interacts with all PMCA b-splice forms. PISP was found to be ubiquitously expressed and, in MDCK cells, was present in a punctate pattern throughout the cytosol and at the basolateral membrane. When added to microsomal membranes expressing PMCA4b, PISP was unable to stimulate the PMCA-dependent ATPase activity. Our data suggest that PISP is a transiently interacting partner of the PMCA b-splice forms that may play a role in their sorting to or from the plasma membrane.

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