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Quality not Quantity: The Role of Marine Natural Products in Drug Discovery and Reverse Chemical Proteomics

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ABSTRACT

Reverse chemical proteomics combines affinity chromatography with phage display and promises to be a powerful new platform technology for the isolation of natural product receptors, facilitating the drug discovery process by rapidly linking biologically active small molecules to their cellular receptors and the receptors’ genes. In this paper we review chemical proteomics and reverse chemical proteomics and show how these techniques can add value to natural products research. We also report on techniques for the derivatisation of polystyrene microtitre plates with cleavable linkers and marine natural products that can be used in chemical proteomics or reverse chemical proteomics. Specifically, we have derivatised polystyrene with palau’amine and used reverse chemical proteomics to try and isolate the human receptors for this potent anticancer marine drug.

No MeSH data available.


Chemical proteomics probes [5] are composed of a tag (or anchor such as polystyrene), a linker (spacer) and probe or reactive group. The probe can be a drug, natural product, peptide, reactive group or anything and can react covalently or non-covalently with its biological receptor, which can have more or less specificity for the probe or react with a specific type of enzyme.
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f2-marinedrugs-03-00036: Chemical proteomics probes [5] are composed of a tag (or anchor such as polystyrene), a linker (spacer) and probe or reactive group. The probe can be a drug, natural product, peptide, reactive group or anything and can react covalently or non-covalently with its biological receptor, which can have more or less specificity for the probe or react with a specific type of enzyme.

Mentions: Chemical proteomics is a powerful tool for isolating and identifying cellular receptors for biologically active natural products, thereby facilitating subsequent rational drug design, and often providing valuable information regarding underlying biochemical and cellular processes. The key to chemical proteomics is the construction of an affinity probe. These probes are composed of three domains (Fig. 2). The tag can consist of either a radioactive/fluorescent label, to allow visualisation of bound proteins on an electrophoresis gel, or a solid-phase bead/surface, to allow affinity purification of proteins. Frequently, biotin is used as the tag as it allows both affinity purification, using a streptavidin resin, and in-gel visualisation, using streptavidin coupled to a reporter enzyme such as horseradish peroxidase. The probe can be any small molecule that reacts with a particular class of enzyme (activity probe) or binds to a particular protein or class of proteins (affinity probe). Affinity probes can be more or less specific for a particular protein or class of proteins or could be completely non-directed [4]. The probe and tag are usually separated from each other by a linker (Fig. 2), that can be simply an alkyl group, peptidic or polyethylene-glycol (PEG) based. The PEG based linkers are hydrophilic like the peptidic linkers, but without the disadvantage of possible cleavage in biological systems. The linker should also be as long as possible as the receptor for a particular small molecule may be deeply buried in the protein.


Quality not Quantity: The Role of Marine Natural Products in Drug Discovery and Reverse Chemical Proteomics
Chemical proteomics probes [5] are composed of a tag (or anchor such as polystyrene), a linker (spacer) and probe or reactive group. The probe can be a drug, natural product, peptide, reactive group or anything and can react covalently or non-covalently with its biological receptor, which can have more or less specificity for the probe or react with a specific type of enzyme.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3756327&req=5

f2-marinedrugs-03-00036: Chemical proteomics probes [5] are composed of a tag (or anchor such as polystyrene), a linker (spacer) and probe or reactive group. The probe can be a drug, natural product, peptide, reactive group or anything and can react covalently or non-covalently with its biological receptor, which can have more or less specificity for the probe or react with a specific type of enzyme.
Mentions: Chemical proteomics is a powerful tool for isolating and identifying cellular receptors for biologically active natural products, thereby facilitating subsequent rational drug design, and often providing valuable information regarding underlying biochemical and cellular processes. The key to chemical proteomics is the construction of an affinity probe. These probes are composed of three domains (Fig. 2). The tag can consist of either a radioactive/fluorescent label, to allow visualisation of bound proteins on an electrophoresis gel, or a solid-phase bead/surface, to allow affinity purification of proteins. Frequently, biotin is used as the tag as it allows both affinity purification, using a streptavidin resin, and in-gel visualisation, using streptavidin coupled to a reporter enzyme such as horseradish peroxidase. The probe can be any small molecule that reacts with a particular class of enzyme (activity probe) or binds to a particular protein or class of proteins (affinity probe). Affinity probes can be more or less specific for a particular protein or class of proteins or could be completely non-directed [4]. The probe and tag are usually separated from each other by a linker (Fig. 2), that can be simply an alkyl group, peptidic or polyethylene-glycol (PEG) based. The PEG based linkers are hydrophilic like the peptidic linkers, but without the disadvantage of possible cleavage in biological systems. The linker should also be as long as possible as the receptor for a particular small molecule may be deeply buried in the protein.

View Article: PubMed Central

ABSTRACT

Reverse chemical proteomics combines affinity chromatography with phage display and promises to be a powerful new platform technology for the isolation of natural product receptors, facilitating the drug discovery process by rapidly linking biologically active small molecules to their cellular receptors and the receptors’ genes. In this paper we review chemical proteomics and reverse chemical proteomics and show how these techniques can add value to natural products research. We also report on techniques for the derivatisation of polystyrene microtitre plates with cleavable linkers and marine natural products that can be used in chemical proteomics or reverse chemical proteomics. Specifically, we have derivatised polystyrene with palau’amine and used reverse chemical proteomics to try and isolate the human receptors for this potent anticancer marine drug.

No MeSH data available.