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Endogenous ADP-ribosylation of eukaryotic elongation factor 2 and its 32 kDa tryptic fragment 47
Ergen,,Kivanç; Bektas¸,Muhammet; Gökçe,Sina; Nurten,Rüstem.
Eukaryotic elongation factor 2 (eEF-2) can undergo ADP-ribosylation in the absence of diphtheria toxin. The binding of free ADP-ribose and endogenous transferase-dependent ADP-ribosylation were distinct reactions for eEF-2, as indicated by different findings. Incubation of eEF-2 tryptic fragment 32/33 kDa (32F) with NAD was ADP-ribosylated and gave rise to the covalent binding of ADP-ribose to eEF-2. 32F was revealed to be at the C-terminal by Edman degradation sequence analysis. In our study, the elution of 32F from SDS-PAGE was ADP-ribosylated both in the presence and absence of diphtheria toxin. These results suggest that endogenous ADP-ribosylation of 32F might be related to protein synthesis. This modification appears to be important for the cell...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Eukaryotic elongation factor 2; Endogenous ADP-ribosylation; Protein synthesis; Trypsin digestion; Diphtheria toxin.
Ano: 2007 URL: http://www.scielo.org.ar/scielo.php?script=sci_arttext&pid=S0327-95452007000100007
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Expression and purification of the immunogenically active fragment B of the Park Williams 8 Corynebacterium diphtheriae strain toxin 56
Nascimento,D.V.; Lemes,E.M.B.; Queiroz,J.L.S.; Silva Jr.,J.G.; Nascimento,H.J.; Silva,E.D.; Hirata Jr.,R.; Dias,A.A.S.O.; Santos,C.S.; Pereira,G.M.B.; Mattos-Guaraldi,A.L.; Armoa,G.R.G..
The construction of a hexahistidine-tagged version of the B fragment of diphtheria toxin (DTB) represents an important step in the study of the biological properties of DTB because it will permit the production of pure recombinant DTB (rDTB) in less time and with higher yields than currently available. In the present study, the genomic DNA of the Corynebacterium diphtheriae Park Williams 8 (PW8) vaccine strain was used as a template for PCR amplification of the dtb gene. After amplification, the dtb gene was cloned and expressed in competent Escherichia coli M15™ cells using the expression vector pQE-30™. The lysate obtained from transformed E. coli cells containing the rDTB PW8 was clarified by centrifugation and purified by affinity chromatography. The...
Tipo: Info:eu-repo/semantics/article Palavras-chave: Fragment B; Diphtheria toxin; Diphtheria; Dtb gene; E. coli gene expression; Immobilized metal affinity.
Ano: 2010 URL: http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2010000500007
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