The regulation of Na, K-ATPase by PLMS, the phospholemman-like protein from shark. Molecular cloning, sequence, expression, cellular distribution and functional effects of PLMS

YA Mahmmoud, Gordon Cramb, AB Maunsbach, Christopher Paul Cutler, L Mieschke, F Cornelius

Research output: Contribution to journalArticlepeer-review

58 Citations (Scopus)

Abstract

In Na, K-ATPase membrane preparations from shark rectal glands, we have previously identified an FXYD domain-containing protein, phospholemman-like protein from shark, PLMS. This protein was shown to associate and modulate shark Na, K-ATPase activity in vitro. Here we describe the complete coding sequence, expression, and cellular localization of PLMS in the rectal gland of the shark Squalus acanthias. The mature protein contained 74 amino acids, including the N-terminal FXYD motif and a C-terminal protein kinase multisite phosphorylation motif. The sequence is preceded by a 20 amino acid candidate cleavable signal sequence. Immunogold labeling of the Na, K-ATPase alpha-subunit and PLMS showed the presence of alpha and PLMS in the basolateral membranes of the rectal gland cells and suggested their partial colocalization. Furthermore, through controlled proteolysis, the C terminus of PLMS containing the protein kinase phosphorylation domain can be specifically cleaved. Removal of this domain resulted in stimulation of maximal Na, K-ATPase activity, as well as several partial reactions. Both the E-1 similar to P --> E-2-P reaction, which is partially rate-limiting in shark, and the K+ deocclusion reaction, E-2(K) --> E-1, are accelerated. The latter may explain the finding that the apparent Na+ affinity was increased by the specific C-terminal PLMS truncation. Thus, these data are consistent with a model where interaction of the phosphorylation domain of PLMS with the Na, K-ATPase alpha-subunit is important for the modulation of shark Na, K-ATPase activity.

Original languageEnglish
Pages (from-to)37427-37438
Number of pages12
JournalJournal of Biological Chemistry
Volume278
Issue number39
DOIs
Publication statusPublished - 26 Sept 2003

Keywords

  • SARCOPLASMIC-RETICULUM CA2+-ATPASE
  • NA+,K+-ATPASE PUMP CYCLE
  • C-TERMINAL DOMAIN
  • GAMMA-SUBUNIT
  • SQUALUS-ACANTHIAS
  • RENAL NA,K-ATPASE
  • RECTAL GLAND
  • KINASE-C
  • INHIBITORY FUNCTION
  • CHANNEL ACTIVITY

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