NDLI: A chemical proteomics based enrichment technique targeting the interactome of the PDE5 inhibitor PF-4540124

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Molecular BioSystems Vol - 5 Issue - 4

Molecular BioSystems Vol - 5 Issue - 3

Molecular BioSystems Vol - 5 Issue - 2

Molecular BioSystems Vol -4

Molecular BioSystems Vol -3

Molecular BioSystems Vol -2

A chemical proteomics based enrichment technique targeting the interactome of the PDE5 inhibitor PF-4540124

Content Provider	Royal Society of Chemistry (RSC)
Author	Scholten, Arjen Rumpel, Klaus Dadvar, Poupak O’Flaherty, Martina Heck, Albert J.R.
Copyright Year	2009
Abstract	The starting point for the discovery and development of new drugs is the design of molecules that bind to their target proteins with high specificity. Here we describe a systematic chemical proteomics based approach, whereby we use a novel PDE5 inhibitor as bait in mice lung tissue. The compound N-(6-aminohexyl)-3-(1-ethyl-3-methyl-7-oxo-6,7-dihydro-1H-pyrazolo[4,3-d]pyrimidin-5-yl)-4-propoxybenzenesulfonamide (or PF-4540124), which binds to phosphodiesterase-5 (PDE5) with high affinity, was therefore immobilized on an affinity support. Initial affinity enrichment revealed the binding of hundreds of proteins to this immobilized PDE5 inhibitor. Therefore, selective pre-clearing and elution protocols were designed and used in combination with differential stable-isotope labeling to discriminate between the specific binding of low abundant proteins and less specific binding of high abundant proteins. The optimized method allowed us to selectively analyze the “interactome” of the PDE5 inhibitorPF-4540124 and enabled us to identify different isoforms of PDE5 present in mouse lung. Additionally, we enriched for the prenyl binding protein PrBP, which is also known as PDE6δ. Further analysis, applying in vitro binding assays allowed us to verify PrBP as a novel interactor of PF-4540124. The presented method provides a generic highly-specific chemical proteomics based enrichment technique for analyzing drug–protein interactions in mammalian tissue lysates.
Starting Page	472
Ending Page	482
Page Count	11
File Format	HTM / HTML PDF
ISSN	1742206X
Volume Number	5
Issue Number	5
Journal	Molecular BioSystems
DOI	10.1039/b815709j
Language	English
Publisher	Royal Society of Chemistry
Access Restriction	Open
Subject Keyword	Mouse Interactome Lung Proteomics CGMP-specific phosphodiesterase type 5 PDE5 inhibitor Protein Phosphodiesterase
Content Type	Text
Resource Type	Article
Subject	Molecular Biology Biotechnology

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Developmental changes in lung cgmp phosphodiesterase-5 activity, protein, and message.

Partial Reconstitution of Photoreceptor cGMP Phosphodiesterase Characteristics in cGMP Phosphodiesterase-5

Potential Roles of Conserved Amino Acids in the Catalytic Domain of the cGMP-binding cGMP-specific Phosphodiesterase (PDE5)

Studies of the Molecular Mechanism of Discrimination between cGMP and cAMP in the Allosteric Sites of the cGMP-binding cGMP-specific Phosphodiesterase (PDE5)

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Cardiovascular effects of sildenafil in hypertensive men with erectile dysfunction and different alleles of the type 5 cGMP-specific phosphodiesterase (PDE5)

Expression of an Active, Monomeric Catalytic Domain of the cGMP-binding cGMP-specific Phosphodiesterase (PDE5)

Cardiovascular effects of sildenafil in hypertensive men with erectile dysfunction and different alleles of type 5 cGMP-specific phosphodiesterase (PDE5)

Expression of cGMP-binding cGMP-specific phosphodiesterase (PDE5) in mouse tissues and cell lines using an antibody against the enzyme amino-terminal domain.

A chemical proteomics based enrichment technique targeting the interactome of the PDE5 inhibitor PF-4540124

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