| RRC ID |
29076
|
| Author |
Bader S, Kortholt A, Snippe H, Van Haastert PJ.
|
| Title |
DdPDE4, a novel cAMP-specific phosphodiesterase at the surface of dictyostelium cells.
|
| Journal |
J Biol Chem
|
| Abstract |
Dictyostelium discoideum cells possess multiple cyclic nucleotide phosphodiesterases that belong either to class I enzymes that are present in all eukaryotes or to the rare beta-lactamase class II. We describe here the identification and characterization of DdPDE4, the third class I enzyme of Dictyostelium. The deduced amino acid sequence predicts that DdPDE4 has a leader sequence, two transmembrane segments, and an extracellular catalytic domain that exhibits a high degree of homology with human cAMP-specific PDE8. Expression of the catalytic domain of DdPDE4 shows that the enzyme is a cAMP-specific phosphodiesterase with a K(m) of 10 microm; cGMP is hydrolyzed at least 100-fold more slowly. The full-length protein is shown to be membrane-bound with catalytic activity exposed to the extracellular medium. Northern blots and activity measurements reveal that expression of DdPDE4 is low during single cell stages and increases at 9 h of starvation, corresponding with mound stage. A function during multicellular development is confirmed by the phenotype of ddpde4(-) knock-out strains, showing normal aggregation but impaired development from the mound stage on. These results demonstrate that DdPDE4 is a unique membrane-bound phosphodiesterase with an extracellular catalytic domain regulating intercellular cAMP during multicellular development.
|
| Volume |
281(29)
|
| Pages |
20018-26
|
| Published |
2006-7-21
|
| DOI |
10.1074/jbc.M600040200
|
| PII |
S0021-9258(19)76316-9
|
| PMID |
16644729
|
| MeSH |
3',5'-Cyclic-AMP Phosphodiesterases / chemistry
3',5'-Cyclic-AMP Phosphodiesterases / genetics*
3',5'-Cyclic-AMP Phosphodiesterases / metabolism*
Amino Acid Sequence
Animals
Cell Membrane / enzymology*
Conserved Sequence
Cyclic AMP
Cyclic Nucleotide Phosphodiesterases, Type 4
DNA Primers
Dictyostelium / enzymology*
Dictyostelium / genetics
Gene Deletion
Humans
Molecular Sequence Data
Phenotype
Polymerase Chain Reaction
Protozoan Proteins / metabolism
Sequence Alignment
Sequence Homology, Amino Acid
|
| IF |
4.238
|
| Times Cited |
16
|
|
WOS Category
|
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
| Resource |
| Cellular slime molds |
S00336 |
