Glutathione fusion proteins

Figure 7-12. Use of glutathione S-transferase (GST) fusion proteins to purify recombinant proteins. (GSH, glutathione.)...

Figure 7.6. Purification of protein from pooled yeast strains. Each yeast ORF was cloned as a fusion to glutathione-S-transferase in a protein expression vector to create 6144 yeast strains. The individual strains were pooled in groups of 96 to create a set of 64 pools. Each pool was grown and the 96 fusion proteins are purified in batch. Each pool was then assayed for a biochemical function (Martzen et al., 1999). Pools positive for function were then deconvoluted using smaller pools consisting of strains from rows and columns of a 96-well plate.

With data obtained by the analysis of fusion proteins consisting of a domain unrelated to poly(3HB) metabolism (e.g., maltose binding protein MalE or glutathione-S-transferase and the poly(3HB) depolymerase binding domain [57,59-61]. 

Shimada K, Nagano M, Kawai M, Koga H. (2005) Influences of amino acid features of glutathione S-transferase fusion proteins on their solubility. Proteomies 5, 3859-63. 

Murray AM, Kelly CD, Nussey SS, Johnstone AP. (1998) Production of glutathione-coated microtitre plates for capturing recombinant glutathione S-transferase fusion proteins as antigens in immunoassays. J Immunol Methods 218, 133-9. 

Rabin DU, Pakner-Crocker R, Mierz DV, Yeung KK. (1992) An ELISA sandwich capture assay for recombinant fusion proteins containing glutathione-S-transferase. J Immunol Methods >6, 101-5. 

Waterboer, T., et al. (2005) Multiplex hiunan papillomavirus serology based on in situ-purified glutathione s-transferase fusion proteins. Clin Chem. 51, 1845-53. 

E)-5-(2-bromovinyl)-2 -deoxyuridine S -monophosphate <1> (<1>, IC50 0.0026 mM, reaction with thymidine, fusion protein with glutathione S-trans-ferase [5]) [5]... 

Problems may be encountered when using recombinant fusion proteins (with P-galactosidase or glutathione transferase) as immunogens, because the response is directed predominantly against the bactenal enzyme. In this case, it is advisable where possible to cleave off and isolate the specific protein... 

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