Isolation and characterization of cDNAs corresponding to two human calcium, calmodulin-regulated, 3′,5′-cyclic nucleotide phosphodiesterase

Loughney, K., Martins, T. J., Harris, E. A. S., Sadhu, K., Hicks, J. B., Sonnenburg, W. K., Beavo, J. A., Ferguson, K. (1996) Isolation and characterization of cDNAs corresponding to two human calcium, calmodulin-regulated, 3′,5′-cyclic nucleotide phosphodiesterase. Journal of Biological Chemistry, 271 (2). pp. 796-806. ISSN 00219258 (ISSN)

Abstract

cDNAs corresponding to two human calcium, calmodulin (CaM)-regulated 3′,5′-cyclic nucleotide phosphodiesterases (PDEs) were isolated. One, Hcam1 (PDE1A3), corresponds to the bovine 61-kDa CaM PDE (PDE1A2). The second, Hcam3 (PDE1C), represents a novel phosphodiesterase gene. Hcaml encodes a 535-amino acid protein that differs most notably from the bovine 61-kDa CaM PDE by the presence of a 14-amino acid insertion and a divergent carboxyl terminus. RNase protection studies indicated that Hcam1 is represented in human RNA from several tissues, including brain, kidney, testes, and heart. Two carboxyl-terminal splice variants for Hcam3 were isolated. One, Hcam3b (PDE1C1), encodes a protein 634 amino acids (72 kDa) in length. The other, Hcam3a (PDE1C3), diverges from Hcam3b 4 amino acids from the carboxyl terminus of Hcam3b, and extends an additional 79 amino acids. All the cDNAs isolated for Hcam3a are incomplete; they do not include the 5′-end of the open reading frame. Northern analysis revealed that both splice variants were expressed in several tissues, including brain and heart, and that there may be additional splice variants. Amino-truncated recombinant proteins were expressed in yeast and characterized biochemically. Hcam3a has a high affinity for both cAMP and cGMP and thus has distinctly different kinetic parameters from Hcam1, which has a higher affinity for cGMP than for cAMP. Both PDE1C enzymes were inhibited by isobutylmethylxanthine, 8-methoxymethyl isobutylmethylxanthine, zaprinast, and vinpocetine.

Item Type: Paper
Uncontrolled Keywords: 3',5' cyclic nucleotide phosphodiesterase calcium calmodulin complementary dna alternative rna splicing amino acid sequence animal cell article binding affinity calcium transport carboxy terminal sequence enzyme binding enzyme isolation nonhuman priority journal second messenger sequence homology signal transduction tissue distribution 3',5'-Cyclic-Nucleotide Phosphodiesterase Animals Base Sequence Cattle Cloning, Molecular DNA, Complementary Humans Molecular Sequence Data Recombinant Proteins Sequence Alignment Sequence Analysis
Subjects: bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification
bioinformatics > genomics and proteomics > genetics & nucleic acid processing
bioinformatics > genomics and proteomics > genetics & nucleic acid processing > DNA, RNA structure, function, modification > cDNA
organs, tissues, organelles, cell types and functions > sub-cellular tissues: types and functions > calcium channel
CSHL Authors:
Communities: CSHL labs > Hicks lab
Depositing User: Matt Covey
Date: 1996
Date Deposited: 09 Aug 2013 20:56
Last Modified: 12 Aug 2013 14:44
Related URLs:
URI: https://repository.cshl.edu/id/eprint/28506

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