Identification of cyclic AMP‐phosphodiesterase variants from the PDE4D gene expressed in human peripheral mononuclear cells

FEBS Letters

Sette, Claudio¹  Zhang, RuoBo¹  Conti, Marco¹  Némoz, Georges¹ 

[1] Division of Reproductive Biology, Department of Gynecology and Obstetrics, Stanford University School of Medicine, Stanford, CA 94305, USA

关键词: Cyclic AMP;    cAMP-phosphodiesterase;    PDE4D gene;    Human lymphocyte;    PDE;    cyclic nucleotide phosphodiesterase;    PKA;    cAMP-dependent protein kinase;    RT-PCR;    reverse transcriptasepolymerase chain reaction;    SDS-PAGE;    polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate;   

DOI  :  10.1016/0014-5793(96)00300-6

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

To determine whether the expression of different PDE4D variants is unique to the rat or conserved through evolution, we have characterized the different PDE4D mRNAs expressed in human peripheral blood mononuclear cells. RT-PCR was performed using primers based on rat sequences and mRNAs from mononuclear cells. The specifically amplified fragments had a size identical to that predicted for rat PDE4D1, PDE4D2 and PDE4D3. Sequencing confirmed that these fragments are derived from the human PDE4D gene. Their sequence was highly homologous to that reported for the rat variants. cDNAs corresponding to the entire ORF of human PDE4D2 and PDE4D3 were expressed in mammalian cells, causing a large increase in PDE activity. Western blot analysis of human peripheral blood mononuclear cell extracts demonstrated the presence of proteins corresponding to the recombinant PDE4D1 and PDE4D2. The pattern of splicing and different promoter usage of the PDE4D gene is therefore conserved during evolution, which indicates an important physiological role.

【 授权许可】

Unknown