Transferases purified

Foureman GL, Hernandez O, Bhatia A, et al. The stereoselectivity of four hepatic glutathione S-transferases purified from a marine elasmobranch (Raja erinacea) with several K-region polycyclic arene oxide substrates. Biochim Biophys Acta 1987 914(2) 127-135. 

ADP-Ribosylation of human c-Ha-ras protein by hen liver ADP-ribosyl transferase. Human c-Ha-ras protein, normal or mutated from glycine to valine at the 12th position, was produced in Escherichia coli and purified (6). These proteins were incubated with ADP-ribosyl transferase purified from hen liver nuclei (7) in the presence of [adenylate- P]NAD. Incorporation of the radioactivity derived from p PJNAD was clearly shown at the positions corresponding to the protein bands of normal and mutated c-Ha-m.y proteins (Fig. lA and IB, lanes 3 and 4). However no significant radioactive band was formed in the absence of c-Ha-ra proteins (Fig. IB, lane 5). The incorporation of the radioactivity into c-Ha-ra.y protein was inhibited by more than 50% in the presence of 40 mM arginine methylester or 40 mM nicotinamide. 

The transferase purified from P. shermanii had a much higher activity than those from P. pentosaceum or Micrococcus lactilicus (Allen et al., 1964). The pH optimum ranged from 6.5 to 7.8 the apparent Km for succinyl-CoA in the transfer to acetate was 1.3 -10 M and for the transfer to propionate it was 6.8 10" M. The enzyme is very stable as evidenced by a... 

ADP-Ribosyl transferase (from human placenta) [9026-30-6]. Purified by making an affinity absorbent for ADP-ribosyltransferase by coupling 3-aminobenzamide to Sepharose 4B. [Burtscher et al. Anal Biochem 152 285 1986.]... 

Soffers AEMF, Ploeman JHTM, Moonen MJH, Wobbes T, van Ommen B, Vervoort J, et al. Regioselectivity and quantitative structure-activity relationships for the conjugation of a series of fluoronitrobenzenes by purified glutathione S -transferase enzymes from rat and man. Chem Res Toxicol 1996 9 638-46. 

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.

In combination of this polymerase with purified propionyl-CoA transferase of Clostridium propionicum, a two-enzyme in vitro PHB biosynthesis system was established which allowed the PHB synthesis from (R)-hydroxybutyric acid as substrate [119]. In this way, the PHB synthesis was independent of the consumption of the expensive CoA, and hence PHA could be readily produced in a semipreparative-scale... 

The exceptional reactivity of aflatoxin B1 exo-8,9-epoxide raises the question of its potential detoxification by EHs. Despite the short half-life, the epoxide does react with DNA (toxification) and glutathione 5-transferases (detoxification), but a role for EH appeared dubious [207], Rat liver or recombinant rat EH has since been shown to provide a modest enhancement of up to 22% in the hydrolysis rate of aflatoxin B1 exo-S,9-epoxide, and to decrease somewhat the genotoxicity of aflatoxin B1 when the ratio of EH to cytochrome P450 is high (ca. 50-fold). Purified human EH provided no such enhancement in hydrolysis, nor did it have a clear effect on genotoxicity. Thus, little evidence exists to support a role for EH in the detoxification of aflatoxin B1 [208],... 

Collins, D. C., Williamson, D. G., and Layne, D. S., Enzymatic synthesis by a partially purified transferase from rabbit liver microsomes. J. Biol. Chem. 245, 873-876 (1970). 

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

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