Alternative splicing generates a CaM kinase IIbeta isoform in myocardium that targets the sarcoplasmic reticulum through a putative alphaKAP and regulates GAPDH

Puneet Singh; John J Leddy; George J Chatzis; Maysoon Salih; Balwant S Tuana

doi:10.1007/s11010-005-5234-y

Alternative splicing generates a CaM kinase IIbeta isoform in myocardium that targets the sarcoplasmic reticulum through a putative alphaKAP and regulates GAPDH

Mol Cell Biochem. 2005 Feb;270(1-2):215-21. doi: 10.1007/s11010-005-5234-y.

Authors

Puneet Singh¹, John J Leddy, George J Chatzis, Maysoon Salih, Balwant S Tuana

Affiliation

¹ Department of Cellular and Molecular Medicine, University of Ottawa, Ottawa, Ontario, Canada.

PMID: 15792370
DOI: 10.1007/s11010-005-5234-y

Abstract

We report the isolation of a full length cDNA from cardiac muscle that encodes a approximately 73 kDa calcium/calmodulin (CaM) dependent kinase IIbeta isoform (CaMKIIbeta(C)) that was generated by alternative splicing of the CaMKIIbeta gene. Antipeptide antibodies raised to specific regions of the kinase identified a 73 kDa kinase polypeptide in cardiac SR. Anti-alpha kinase anchoring protein (alphaKAP) antibodies identified a 25 kDa polypeptide in cardiac SR and RT-PCR followed by sequence analysis confirmed the presence of a full length alphaKAP encoding transcript in myocardium. Protein interaction assays revealed that the 73 kDa CaMKIIbeta(C) binds GAPDH to modulate the production of NADH in a Ca2+/CaM dependent reaction. The presence of a CaMKIIbeta isoform that can target the SR presumably via its membrane anchor alphaKAP defines a previously unrecognized Ca2+/CaM regulatory system in myocardium.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Alternative Splicing*
Amino Acid Sequence
Animals
Base Sequence
Blotting, Northern
Calcium-Calmodulin-Dependent Protein Kinase Type 2
Calcium-Calmodulin-Dependent Protein Kinases / chemistry*
Calcium-Calmodulin-Dependent Protein Kinases / genetics*
Cells, Cultured
Cytosol / metabolism
DNA, Complementary / metabolism
Electrophoresis, Polyacrylamide Gel
Exons
Gene Library
Glutathione Transferase / metabolism
Glyceraldehyde-3-Phosphate Dehydrogenases / biosynthesis*
Humans
Immunoblotting
Introns
Molecular Sequence Data
Muscle, Skeletal / metabolism
Myocardium / enzymology
Myocardium / metabolism*
Myocardium / pathology*
NAD / metabolism
Peptides / chemistry
Protein Binding
Protein Isoforms
Protein Structure, Tertiary
Rabbits
Sarcoplasmic Reticulum / metabolism
Sequence Homology, Amino Acid
Tissue Distribution

Substances

DNA, Complementary
Peptides
Protein Isoforms
NAD
Glyceraldehyde-3-Phosphate Dehydrogenases
Glutathione Transferase
CAMK2B protein, human
Calcium-Calmodulin-Dependent Protein Kinase Type 2
Calcium-Calmodulin-Dependent Protein Kinases