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Inositol 1,4,5-trisphosphate receptor localization and stability in neonatal cardiomyocytes requires interaction with ankyrin-B.

Publication ,  Journal Article
Mohler, PJ; Davis, JQ; Davis, LH; Hoffman, JA; Michaely, P; Bennett, V
Published in: J Biol Chem
March 26, 2004

The molecular mechanisms required for inositol 1,4,5-trisphosphate receptor (InsP(3)R) targeting to specialized endoplasmic reticulum membrane domains are unknown. We report here a direct, high affinity interaction between InsP(3)R and ankyrin-B and demonstrate that this association is critical for InsP(3)R post-translational stability and localization in cultures of neonatal cardiomyocytes. Recombinant ankyrin-B membrane-binding domain directly interacts with purified cerebellar InsP(3)R (K(d) = 2 nm). 220-kDa ankyrin-B co-immunoprecipitates with InsP(3)R in tissue extracts from brain, heart, and lung. Alanine-scanning mutagenesis of the ankyrin-B ANK (ankyrin repeat) repeat beta-hairpin loop tips revealed that consecutive ANK repeat beta-hairpin loop tips (repeats 22-24) are required for InsP(3)R interaction, thus providing the first detailed evidence of how ankyrin polypeptides associate with membrane proteins. Pulse-chase biosynthesis experiments demonstrate that reduction or loss of ankyrin-B in ankyrin-B (+/-) or ankyrin-B (-/-) neonatal cardiomyocytes leads to approximately 3-fold reduction in half-life of newly synthesized InsP(3)R. Furthermore, interactions with ankyrin-B are required for InsP(3)R stability as abnormal InsP(3)R phenotypes, including mis-localization, and reduced half-life in ankyrin-B (+/-) cardiomyocytes can be rescued by green fluorescent protein (GFP)-220-kDa ankyrin-B but not by GFP-220-kDa ankyrin-B mutants, which do not associate with InsP(3)R. These new results provide the first physiological evidence of a molecular partner required for early post-translational stability of InsP(3)R.

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Published In

J Biol Chem

DOI

ISSN

0021-9258

Publication Date

March 26, 2004

Volume

279

Issue

13

Start / End Page

12980 / 12987

Location

United States

Related Subject Headings

  • Tissue Distribution
  • Time Factors
  • Recombinant Proteins
  • Receptors, Cytoplasmic and Nuclear
  • Protein Structure, Tertiary
  • Protein Structure, Secondary
  • Protein Processing, Post-Translational
  • Protein Binding
  • Precipitin Tests
  • Phenotype
 

Citation

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Mohler, P. J., Davis, J. Q., Davis, L. H., Hoffman, J. A., Michaely, P., & Bennett, V. (2004). Inositol 1,4,5-trisphosphate receptor localization and stability in neonatal cardiomyocytes requires interaction with ankyrin-B. J Biol Chem, 279(13), 12980–12987. https://doi.org/10.1074/jbc.M313979200
Mohler, Peter J., Jonathan Q. Davis, Lydia H. Davis, Janis A. Hoffman, Peter Michaely, and Vann Bennett. “Inositol 1,4,5-trisphosphate receptor localization and stability in neonatal cardiomyocytes requires interaction with ankyrin-B.J Biol Chem 279, no. 13 (March 26, 2004): 12980–87. https://doi.org/10.1074/jbc.M313979200.
Mohler PJ, Davis JQ, Davis LH, Hoffman JA, Michaely P, Bennett V. Inositol 1,4,5-trisphosphate receptor localization and stability in neonatal cardiomyocytes requires interaction with ankyrin-B. J Biol Chem. 2004 Mar 26;279(13):12980–7.
Mohler, Peter J., et al. “Inositol 1,4,5-trisphosphate receptor localization and stability in neonatal cardiomyocytes requires interaction with ankyrin-B.J Biol Chem, vol. 279, no. 13, Mar. 2004, pp. 12980–87. Pubmed, doi:10.1074/jbc.M313979200.
Mohler PJ, Davis JQ, Davis LH, Hoffman JA, Michaely P, Bennett V. Inositol 1,4,5-trisphosphate receptor localization and stability in neonatal cardiomyocytes requires interaction with ankyrin-B. J Biol Chem. 2004 Mar 26;279(13):12980–12987.

Published In

J Biol Chem

DOI

ISSN

0021-9258

Publication Date

March 26, 2004

Volume

279

Issue

13

Start / End Page

12980 / 12987

Location

United States

Related Subject Headings

  • Tissue Distribution
  • Time Factors
  • Recombinant Proteins
  • Receptors, Cytoplasmic and Nuclear
  • Protein Structure, Tertiary
  • Protein Structure, Secondary
  • Protein Processing, Post-Translational
  • Protein Binding
  • Precipitin Tests
  • Phenotype