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High-throughput screening for transglutaminase activities using recombinant fluorescent proteins

, : High-throughput screening for transglutaminase activities using recombinant fluorescent proteins. Biotechnology and Bioengineering 110(11): 2865-2873

Since detailed evaluation of specific transglutaminases (TGs) from various species requires identification of their substrate specificities, rapid substrate screening method by measurement of their relative activities is in great demand. Here, a novel evaluation method of TG activity was developed using two recombinant fluorescent proteins (FPs), that is, eYFP and DsRed, tagged with TG substrate peptides. By cross-linking the two FPs based on the tagged target peptide sequences at their C-terminus, the expression of co-transformed TG allows quenching of the yellow fluorescence intensities. It was shown that the degree of in vivo fluorescent quenching by the TG activity agrees well with its in vitro reaction data, suggesting that this system can be used to identify relative substrate specificity of TGs for target peptide sequences. Using this method, the lysine substrates of TGs from Bacillus species (BTG) were evaluated, and the newly selected pentapeptide, KTKTN showed almost the same reactivity with the well-known hexa-lysine (K₆) substrate for BTG reaction.


PMID: 23740563

DOI: 10.1002/bit.24970

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Raymond, E.; Louvet, C.; Tournigand, C.; Coudray, A.Marie.; Faivre, S.; D.G.amont, A.; Gespach, C., 2002: Pemetrexed disodium combined with oxaliplatin, SN38, or 5-fluorouracil, based on the quantitation of drug interactions in human HT29 colon cancer cells. Premetrexed disodium (MTA) is a novel multi-targeted antifolate that inhibits thymidylate synthase, dihydrofolate reductase, and glycinamide ribonucleotide formyltransferase. It exhibits a broad spectrum of activity against several human tumor typ...

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Hoagland, T.A.; Jeong, B.S.on; Riesen, J.W.; Schreiber, D.; Yang, X.; Fodor, W.L., 1999: The yield of ova from the ovaries of 20, 60, and 115 kg gilts treated with or without PMSG. Journal of Animal Science 77(SUPPL 1): 92

Grafton-Cardwell, E.E.; Eash, J.A.; Granett, J., 1988: Isozyme differentiation of Tetranychus pacificus from T urticae and T turkestani Acari Tetranychidae in laboratory and field populations. Cellulose acetate electrophoresis distinguished among females of three spider mite species, Tetranychus turkestani Ugarov & Nikolski, T. urticae Koch, and T. pacificus McGregor. Phosphoglucose isomerase and malate dehydrogenase separated both labo...

Kazarinov, B.N., 1972: Weight gains of young cattle in relation to the soluble fractions of protein in the feed. Kholmogor bulls 6 to 7 months old were given diets of about the same energy value and based on grass hay, rolled oats barley and peas, linseed cake, grass meal, salt and CaCO3. Readily soluble proteins accounted for 49, 40 or 32.1% of total protei...

Olofsson, O., 1934: Ornitologiskt strandhugg i Norrbottens skargard. Fauna och Flora Uppsala, 173-176

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Pernar, N.; Baksic, D.; Antonic, O.; Grubesic, M.; Tikvic, I.; Trupcevic, M., 2006: Oil residuals in lowland forest soil after pollution with crude oil. Research on oil residuals in lowland forest soil was carried out in 6 sample plots in the lowland forest ecosystem located in an oil field. Four plots were differently affected in terms of discharged oil and the time lapsed after the accident, as...

Gamon, Ja, 1987: Advisory councils--real friends. Journal of extension 25(25): 42-43