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

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.


Agarose gel electrophoresis of PCR products obtained from various phage-displayed human cDNA libraries after five rounds of selection with palau’amine immobilised on a PS plate via a cleavable disulfide-containing linker. The DNA inserts were amplified using generic T7 primers.
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f12-marinedrugs-03-00036: Agarose gel electrophoresis of PCR products obtained from various phage-displayed human cDNA libraries after five rounds of selection with palau’amine immobilised on a PS plate via a cleavable disulfide-containing linker. The DNA inserts were amplified using generic T7 primers.

Mentions: Palau’amine was immobilised on a bromomethylated PS microtitre plate via a cleavable disulfide linker and the resulting affinity support was used to probe five different T7 phage-displayed human cDNA libraries. Following each round of selection, phages retained by the affinity support were released by reductive cleavage of the disulfide linker with DTT. After five rounds of selection had been performed, 16 individual plaques were picked from each sublibrary and their DNA inserts were amplified by PCR using generic T7 primers (Fig. 13). In addition, a subset of the amplified DNA inserts from each sublibrary was sequenced (Table 1).


Quality not Quantity: The Role of Marine Natural Products in Drug Discovery and Reverse Chemical Proteomics
Agarose gel electrophoresis of PCR products obtained from various phage-displayed human cDNA libraries after five rounds of selection with palau’amine immobilised on a PS plate via a cleavable disulfide-containing linker. The DNA inserts were amplified using generic T7 primers.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3756327&req=5

f12-marinedrugs-03-00036: Agarose gel electrophoresis of PCR products obtained from various phage-displayed human cDNA libraries after five rounds of selection with palau’amine immobilised on a PS plate via a cleavable disulfide-containing linker. The DNA inserts were amplified using generic T7 primers.
Mentions: Palau’amine was immobilised on a bromomethylated PS microtitre plate via a cleavable disulfide linker and the resulting affinity support was used to probe five different T7 phage-displayed human cDNA libraries. Following each round of selection, phages retained by the affinity support were released by reductive cleavage of the disulfide linker with DTT. After five rounds of selection had been performed, 16 individual plaques were picked from each sublibrary and their DNA inserts were amplified by PCR using generic T7 primers (Fig. 13). In addition, a subset of the amplified DNA inserts from each sublibrary was sequenced (Table 1).

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.