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Comparative analysis of depurination catalyzed by ricin A-chain on synthetic 32mer and 25mer oligoribonucleotides mimicking the sarcin/ricin domain of the rat 28S rRNA and E. coli 23S rRNA

Paper ID Volume ID Publish Year Pages File Format Full-Text
24886 43543 2009 7 PDF Available
Title
Comparative analysis of depurination catalyzed by ricin A-chain on synthetic 32mer and 25mer oligoribonucleotides mimicking the sarcin/ricin domain of the rat 28S rRNA and E. coli 23S rRNA
Abstract

Ricin A-chain can inactivate eukaryotic ribosomes, but exhibits no N-glycosidase activity on intact E. coli ribosomes. In the present research, in order to avoid using radiolabeled oligoribonucleotides, two kinds of synthetic 5′-FAM fluorescence-labeled oligoribonucleotide substrates were used to mimic the sarcin/ricin domains of rat 28S rRNA and E. coli 23S rRNA (32mer and 25mer, named as Rat FAM-SRD and E. coli FAM-SRD, respectively). Ricin A-chain was able to specifically release adenine from the first adenosine of the GAGA tetraloop and exhibited specific N-glycosidase activity under neutral and weak acidic conditions with both substrates. However, under more acidic conditions, ricin A-chain was able to release purines from other sites on eukaryotic substrates, but it retained specific depurination activity on prokaryotic substrates. At pH 5.0, the Michaelis constant (Km) for the reaction with Rat FAM-SRD (4.57 ± 0.28 μM) corresponded to that with E. coli FAM-SRD (4.64 ± 0.26 μM). However, the maximum velocity (Vmax) for ricin A-chain with Rat FAM-SRD was 0.5 ± 0.024 μM/min, which is higher than that with E. coli FAM-SRD (0.32 ± 0.011 μM/min).

Keywords
RIPs, ribosome-inactivating proteins; RTA, ricin A-chain; SRD, sarcin/ricin domain RNA; Rat FAM-SRD, synthetic 32mer 5′-FAM fluorescence-labeled oligoribonucleotide mimicking the sarcin/ricin domains of rat 28S rRNA; E. coli FAM-SRD, synthetic 25mer 5′-FA
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Comparative analysis of depurination catalyzed by ricin A-chain on synthetic 32mer and 25mer oligoribonucleotides mimicking the sarcin/ricin domain of the rat 28S rRNA and E. coli 23S rRNA
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 139, Issue 2, 15 January 2009, Pages 156–162
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us