Edman degradation and protein sequencing: Pfu Pyroglutamate Aminopeptidase
Pfu Pyroglutamate Aminopeptidase removes pyroglutamic acids from the N-termini of proteins and peptides, enabling protein sequencing following Edman degradation.
This enzyme may work well with some intact and non-denatured proteins; a denaturation step may be unnecessary in some cases. Pfu Pyroglutamate Aminopeptidase is supplied with a 5X Reaction Buffer [250 mM sodium phosphate (pH 7.0), 50 mM DTT, 5 mM EDTA].
More Information
Applications
- Removal of pyroglutamic acids from the N-termini of proteins and peptides
Components
Supplied Buffer
- Volume: 1 ml
- Component: 250 mM sodium phosphate buffer (pH 7.0) containing 50 mM DTT and 5 mM EDTA
- Comparison of specific activities across different temperatures: 5.83 U/mg protein at 37°C showed activities of 12.2 U/mg protein or 28.3 U/mg protein at 50°C or 75°C, respectively.
Source
Recombinant Escherichia coli encoding the Pyrococcus furiosus pyroglutamate aminopeptidase gene.
Properties
| Systematic name: | L-Pyrrolidone carboxyl peptidase |
| Enzyme code: | 3.4.19.3 |
| Molecular weight: | 24.072 kDA (calculated), 28 kDA (by SDS-PAGE) |
| Optimum temperature: | 95–100°C |
| Thermo stability: | ~90% activity at 75°C; pH 7.0, 150 min |
| Optimum pH: | 6.0–9.0; ≥80% activity in a pH range of 5.0–9.0 at 75°C when reactivated for 30 min |
| Tolerance to denaturants: | ≤1 M Urea, ≤1 M Guanidine-HCL, ≤0.01% SDS |
| Inhibitors: | PCMB, Hg2+ |
Definition of activity
One unit of enzyme activity corresponds to the amount required to hydrolyze 1 µmol pyroglutamate p-nitroanilide at 37°C in 1 minute at pH 7.0.
References
Hamazume, Y. & Mega, T. Positions of Disulfide Bonds in Riboflavin-Binding Protein of Hen Egg White. J. Biochem 101, 217–223 (1987).
Shimada, Y., Sugihara, a, Tominaga, Y., Iizumi, T. & Tsunasawa, S. cDNA molecular cloning of Geotrichum candidum lipase. J. Biochem. 106, 383–388 (1989).
Additional product information
Please see the product's Certificate of Analysis for information about storage conditions, product components, and technical specifications. Please see the Kit Components List to determine kit components. Certificates of Analysis and Kit Components Lists are located under the Documents tab.
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