| Autor: |
Amegadzie, Bernard Y., Hanning, Charles R., McLaughlin, Megan M., Burman, Miriam, Cieslinski, Lenora B., Livi, George P., Torphy, Theodore J. |
| Zdroj: |
Cell Biology International; June 1995, Vol. 19 Issue: 6 p477-484, 8p |
| Abstrakt: |
Recombinant baculoviruses were constructed to express cDNAs encoding two distinct subtypes of human cAMP-specific phosphodiesterase (hPDE4A and hPDE4B). Infection of Spodoptera frugiperda insect cells with the appropriate recombinant baculoviruses resulted in high level production of biologically-active protein as measured by enzymatic activity and immunoblotting using subtype-specific anti-hPDE4 antisera. Both recombinant proteins showed catalytic activity with a low Km (~ 3 µM) for cAMP (with no cGMP hydrolyzing activity) and were inhibited by R-rolipram with apparent Kis of 0.38 and 0.25 µM, respectively. The recombinant enzymes also contained saturable, stereoselective and high-affinity rolipram-binding sites (Kd ~ 2 nM). Thus, insect cell-derived hPDE4s possess kinetic properties analogous to native enzymes as well as to recombinant enzymes produced in yeast. Copyright 1995, 1999 Academic Press |
| Databáze: |
Supplemental Index |
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