mRNA interferase-MazF enzyme
MazF enzyme is a toxin protein in the toxin-antitoxin module of E. coli. It possesses endoribonuclease activity and specifically cleaves single-stranded RNA (ssRNA) at 5' ACA sequences. MazF does not cleave double-stranded RNA (dsRNA), double-stranded DNA, or single-stranded DNA. The mRNA interferase-MazF enzyme is supplied as a fusion protein of E. coli MazF and Trigger Factor, which is an E. coli chaperone protein. The enzyme is also supplied with 5X MazF Buffer consisting of 200 mM sodium phosphate (pH 7.5) and 0.05% Tween-20.
- Use for site-dependent cleavage of ssRNA
Expressed as a recombinant protein in Escherichia coli.
Additional MazF nuclease activities were not detected under any of the following conditions as demonstrated by RNA or DNA electrophoresis:
- After incubation with 40 pmol non-AC sequence ssRNA and 20 units mRNA Interferase-MazF for 16 hr at 37°C and pH 7.5.
- After incubation with 40 pmol dsRNA and 20 units mRNA Interferase-MazF for 16 hr at 37°C and pH 7.5.
- After incubation with 1 mg lambda DNA-Hind III fragments and 20 units mRNA Interferase-MazF for 16 hr at 37°C and pH 7.5.
Licensing agreement required. This product is limited to research use only. Product must not be used for clinical or in vitro diagnostic purposes.
Provided as a solution of 20 mM sodium phosphate (pH 6.0), 0.01% Tween-20 and 50% glycerol.
Definition of activity
One unit is defined as the amount of enzyme that cleaves 1 pmol of standard substrate ROX-5'-GATAUACATATCT-eclipse (the underlined region is composed of RNA) after 10 min at 37°C and pH 7.5. Buffer composition used for activity definition: 20 mM sodium phosphate, pH 7.5 and 0.05% Tween-20.
Suzuki, M., Zhang, J., Liu, M., Woychik, N. A. & Inouye, M. Single Protein Production in Living Cells Facilitated by an mRNA Interferase. Mol. Cell 18 (2005).
Zhang, Y. et al. MazF Cleaves Cellular mRNAs Specifically at ACA to Block Protein Synthesis in Escherichia coli. Mol. Cell 12, 913–923 (2003).
Zhang, Y., Zhang, J., Hara, H., Kato, I. & Inouye, M. Insights into the mRNA cleavage mechanism by MazF, an mRNA interferase. J. Biol. Chem. 280, 3143–50 (2005).
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