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Characterization of a protein cofactor that mediates protein kinase A regulation of the renal brush border membrane Na(+)-H+ exchanger.

Publication ,  Journal Article
Weinman, EJ; Steplock, D; Wang, Y; Shenolikar, S
Published in: J Clin Invest
May 1995

Activation of cAMP-dependent protein kinase A inhibits the renal proximal tubule brush border membrane Na(+)-H+ exchanger by a process involving participation of a regulatory cofactor (NHE-RF) that is distinct from the transporter itself. Recent studies from this laboratory reported a partial amino acid sequence of this putative cofactor (Weinman, E. J., D. H. Steplock, and S. Shenolikar. 1993. J. Clin. Invest. 92:1781-1786). The present experiments detail the structure of the NHE-RF protein as determined from molecular cloning studies. A codon-biased oligonucleotide probe to a portion of the amino acid sequence of the putative cofactor was used to isolate a 1.9-kb cDNA from a rabbit renal library. The encoded protein is 358 amino acids in length and is rich in proline residues. Search of existing data bases indicates that NHE-RF is a unique protein. Using a reticulocyte lysate, the cDNA translated a product of approximately 44 kD, which was recognized by an affinity-purified polyclonal antibody to NHE-RF. Potential phosphorylation sites for protein kinase A are present. The mRNA for the protein is expressed in kidney, proximal small intestine, and liver. Reverse transcription/PCR studies in the kidney indicate the presence of mRNA for NHE-RF in several distinct nephron segments including the proximal tubule.

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Published In

J Clin Invest

DOI

ISSN

0021-9738

Publication Date

May 1995

Volume

95

Issue

5

Start / End Page

2143 / 2149

Location

United States

Related Subject Headings

  • Sodium-Hydrogen Exchangers
  • Sequence Homology, Amino Acid
  • Recombinant Proteins
  • Rabbits
  • Protein Conformation
  • Polymerase Chain Reaction
  • Phosphoproteins
  • Organ Specificity
  • Oligonucleotide Probes
  • Molecular Sequence Data
 

Citation

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Weinman, E. J., Steplock, D., Wang, Y., & Shenolikar, S. (1995). Characterization of a protein cofactor that mediates protein kinase A regulation of the renal brush border membrane Na(+)-H+ exchanger. J Clin Invest, 95(5), 2143–2149. https://doi.org/10.1172/JCI117903
Weinman, E. J., D. Steplock, Y. Wang, and S. Shenolikar. “Characterization of a protein cofactor that mediates protein kinase A regulation of the renal brush border membrane Na(+)-H+ exchanger.J Clin Invest 95, no. 5 (May 1995): 2143–49. https://doi.org/10.1172/JCI117903.
Weinman, E. J., et al. “Characterization of a protein cofactor that mediates protein kinase A regulation of the renal brush border membrane Na(+)-H+ exchanger.J Clin Invest, vol. 95, no. 5, May 1995, pp. 2143–49. Pubmed, doi:10.1172/JCI117903.

Published In

J Clin Invest

DOI

ISSN

0021-9738

Publication Date

May 1995

Volume

95

Issue

5

Start / End Page

2143 / 2149

Location

United States

Related Subject Headings

  • Sodium-Hydrogen Exchangers
  • Sequence Homology, Amino Acid
  • Recombinant Proteins
  • Rabbits
  • Protein Conformation
  • Polymerase Chain Reaction
  • Phosphoproteins
  • Organ Specificity
  • Oligonucleotide Probes
  • Molecular Sequence Data