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Identification of a novel isoform of the cyclic-nucleotide phosphodiesterase PDE3A expressed in vascular smooth-muscle myocytes

Choi, Young-Hun; Ekholm, Dag; Krall, Judith; Ahmad, Faiyaz; Degerman, Eva LU ; Manganiello, Vincent C. and Movsesian, Matthew A. (2001) In Biochemical Journal 353(Pt 1). p.41-50
Abstract
We have identified a new cyclic-nucleotide phosphodiesterase isoform, PDE3A, and cloned its cDNA from cultured aortic myocytes. The nucleotide sequence of its coding region is similar to that of the previously cloned myocardial isoform except for the absence of the initial 300-400 nt that are present in the latter, as confirmed by reverse-transcriptase-mediated PCR, 5' rapid amplification of cDNA ends and a ribonuclease protection assay. Expression in Spodoptera frugiperda (Sf9) cells yields a protein with catalytic activity and inhibitor sensitivity typical of the PDE3 family. The recombinant protein's molecular mass of approx. 131 kDa is compatible with translation from an ATG sequence corresponding to nt 436-438 of the myocardial PDE3A... (More)
We have identified a new cyclic-nucleotide phosphodiesterase isoform, PDE3A, and cloned its cDNA from cultured aortic myocytes. The nucleotide sequence of its coding region is similar to that of the previously cloned myocardial isoform except for the absence of the initial 300-400 nt that are present in the latter, as confirmed by reverse-transcriptase-mediated PCR, 5' rapid amplification of cDNA ends and a ribonuclease protection assay. Expression in Spodoptera frugiperda (Sf9) cells yields a protein with catalytic activity and inhibitor sensitivity typical of the PDE3 family. The recombinant protein's molecular mass of approx. 131 kDa is compatible with translation from an ATG sequence corresponding to nt 436-438 of the myocardial PDE3A coding region. Antibodies against residues 424-460 (nt 1270-1380) and 1125-1141 (nt 3373-3423) of the myocardial isoform react with an approx. 118 kDa band in Western blots of homogenates of human aortic myocytes, whereas antibodies against residues 29-42 (nt 85-126) do not react with any bands in these homogenates. Our results suggest that a vascular smooth-muscle isoform ('PDE3A2') is a product of the same gene as the longer myocardial ('PDE3A1') and the shorter placental ('PDE3A3') isoforms and is generated pre-translationally in a manner that results in the absence of the 145 N-terminal amino acids of PDE3A1. (Less)
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author
organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
intracellular signalling, protein phosphorylation, second messengers, vasodilation
in
Biochemical Journal
volume
353
issue
Pt 1
pages
41 - 50
publisher
Portland Press Limited
external identifiers
  • pmid:11115397
  • scopus:0035148960
ISSN
0264-6021
language
English
LU publication?
yes
id
ed537726-b305-4972-b009-5950d1f53f7e (old id 1122457)
date added to LUP
2008-06-26 09:02:16
date last changed
2018-05-29 10:54:52
@article{ed537726-b305-4972-b009-5950d1f53f7e,
  abstract     = {We have identified a new cyclic-nucleotide phosphodiesterase isoform, PDE3A, and cloned its cDNA from cultured aortic myocytes. The nucleotide sequence of its coding region is similar to that of the previously cloned myocardial isoform except for the absence of the initial 300-400 nt that are present in the latter, as confirmed by reverse-transcriptase-mediated PCR, 5' rapid amplification of cDNA ends and a ribonuclease protection assay. Expression in Spodoptera frugiperda (Sf9) cells yields a protein with catalytic activity and inhibitor sensitivity typical of the PDE3 family. The recombinant protein's molecular mass of approx. 131 kDa is compatible with translation from an ATG sequence corresponding to nt 436-438 of the myocardial PDE3A coding region. Antibodies against residues 424-460 (nt 1270-1380) and 1125-1141 (nt 3373-3423) of the myocardial isoform react with an approx. 118 kDa band in Western blots of homogenates of human aortic myocytes, whereas antibodies against residues 29-42 (nt 85-126) do not react with any bands in these homogenates. Our results suggest that a vascular smooth-muscle isoform ('PDE3A2') is a product of the same gene as the longer myocardial ('PDE3A1') and the shorter placental ('PDE3A3') isoforms and is generated pre-translationally in a manner that results in the absence of the 145 N-terminal amino acids of PDE3A1.},
  author       = {Choi, Young-Hun and Ekholm, Dag and Krall, Judith and Ahmad, Faiyaz and Degerman, Eva and Manganiello, Vincent C. and Movsesian, Matthew A.},
  issn         = {0264-6021},
  keyword      = {intracellular signalling,protein phosphorylation,second messengers,vasodilation},
  language     = {eng},
  number       = {Pt 1},
  pages        = {41--50},
  publisher    = {Portland Press Limited},
  series       = {Biochemical Journal},
  title        = {Identification of a novel isoform of the cyclic-nucleotide phosphodiesterase PDE3A expressed in vascular smooth-muscle myocytes},
  volume       = {353},
  year         = {2001},
}