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Cloning and expression of cDNA for a human low-K sub m , rolipram-sensitive cyclic AMP phosphodiesterase

Journal Article · · Molecular and Cellular Biology; (USA)
; ; ;  [1]; ;  [2]; ;  [3];  [4]
  1. Dept. of Gene Expression Sciences, Smithkline Beecham Pharmaceuticals, King of Prussia, PA (US)
  2. Dept. of Molecular Genetics, Smithkline Beecham Pharmaceuticals, King of Prussia, PA (US)
  3. Dept. of Pharmacology, Smithkline Beecham Pharmaceuticals, King of Prussia, PA (US)
  4. Baylor Univ., Houston, TX (USA). Dept. of Cell Biology
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The authors have isolated cDNA clones representing cyclic AMP (cAMP)-specific phosphodiesterases (PDEases) from a human monocyte cDNA library. One cDNA clone (hPDE-1) defines a large open reading frame of ca. 2.1 kilobases, predicting a 686-amino-acid, ca. 77-kilodalton protein which contains significant homology to both rat brain and {ital Drosophila} cAMP PDEases, especially within an internal conserved domain of ca. 270 residues. Amino acid sequence divergence exists at the NH{sub 2} terminus and also within a 40- to 100-residue domain near the COOH-terminal end. hPDE-1 hybridizes to a major 4.8-kilobase mRNA transcript from both human monocytes and placenta. The coding region of hPDE-1 was engineered for expression in COS-1 cells, resulting in the overproduction of cAMP PDEase activity. The hPDE-1 recombinant gene product was identified as a low-{ital K{sub m}} cAMP phosphodiesterase on the basis of several biochemical properties including selective inhibition by the antidepressant drug rolipram. Known inhibitors of other PDEases (cGMP-specific PDEase, cGMP-inhibited PDEase) had little or no effect on the hPDE-1 recombinant gene product.

OSTI ID:
6511588
Journal Information:
Molecular and Cellular Biology; (USA), Journal Name: Molecular and Cellular Biology; (USA) Vol. 10:6; ISSN MCEBD; ISSN 0270-7306
Country of Publication:
United States
Language:
English

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Related Subjects

550200* -- Biochemistry
550400 -- Genetics
59 BASIC BIOLOGICAL SCIENCES
AMINO ACID SEQUENCE
AMP
ANIMALS
ARTHROPODS
BIOLOGICAL FUNCTIONS
BIOLOGICAL MATERIALS
BIOSYNTHESIS
BLOOD
BLOOD CELLS
BODY FLUIDS
CLONING
DIPTERA
DNA
DNA HYBRIDIZATION
DNA-CLONING
DROSOPHILA
ENZYMES
ESTERASES
FLIES
FRUIT FLIES
FUNCTIONS
GENE RECOMBINATION PROTEINS
GENE REGULATION
HYBRIDIZATION
HYDROLASES
INFLAMMATION
INSECTS
INVERTEBRATES
LEUKOCYTES
MATERIALS
MOLECULAR STRUCTURE
MONOCYTES
NUCLEIC ACIDS
NUCLEOPROTEINS
NUCLEOTIDES
ORGANIC COMPOUNDS
PATHOLOGICAL CHANGES
PHENOTYPE
PHOSPHODIESTERASES
PROTEINS
RECOMBINANT DNA
SYMPTOMS
SYNTHESIS