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Production and Characterization of Pharmacologically Active Recombinant Human Phosphodiesterase 4B in Dictyostelium Discoideum

Production and Characterization of Pharmacologically Active Recombinant Human Phosphodiesterase 4B in Dictyostelium Discoideum

Ranjana Arya, Saima Aslam, Shivani Gupta, Roop Singh Bora, Lalitha Vijayakrishnan, Pankaj Gulati, Sudha Naithani, Shohini Mukherjee, Sunanda Dastidar, Alok Bhattacharya, Kulvinder Singh Saini

Biotechnol J. 2008 Jul;3(7):938-47.

PMID: 18348140

Abstract:

Phosphodiesterase 4B (PDE4B) is an important therapeutic target for asthma and chronic obstructive pulmonary disease. To identify PDE4 subtype-specific compounds using high-throughput assays, full-length recombinant PDE4 proteins are needed in bulk quantity. In the present study, full-length human PDE4B2 was expressed in the cellular slime mould Dictyostelium discoideum (Dd). A cell density of 2 x 10(7) cells/mL was obtained and up to 1 mg/L recombinant PDE4B2 was purified through Ni-NTA affinity chromatography. The expressed protein was soluble and its activity was comparable to PDE4B2 protein expressed in mammalian cells (K(m)=1.7 microM). The functional significance of the Dd expression system is supported by the demonstration that, in concert with proteins expressed in mammalian systems, there are no major changes in the affinity for PDE4B2 inhibitors and substrates. These findings thus provide the first evidence that Dd can be utilized for the expression and purification of functionally active full-length human PDE4B2 in large amounts required for high-throughput screening of pharmacologically active compounds against this therapeutic target.

Chemicals Related in the Paper:

Catalog Number	Product Name	Structure	CAS Number	Price
IAR4245702	PDE4B2 Active human			Price

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