Tryp-N™, or “N-terminal trypsin”, is a specialized protease from ProtiFi™ that cleaves proteins at the N-terminal side of lysine or arginine residues.
Tryp-N™ is a thermophilic metalloprotease with N-terminal specificity for arginine and lysine engineered at Cold Spring Harbor Laboratory (CSHL). It provides clarified spectra and greater sensitivity by yielding peptides in which the basic centers of peptides – the amino terminus and side chains of K/R – are located together to produce b-ion dominated spectra. Tryp-N has been demonstrated to yield simplified MS/MS fragmentation and have the best proteolytic activity.¹ Tryp-N spectra provide the complimentary ion series to compliment trypsin, allowing you to more easily find and specifically localize PTMs. PTM identification and localization of PTMs, such as phosphorylation in YST clusters, and de novo sequencing. Additionally, as ion current remains on b-ions during fragmentation of MRM assays, rather than becoming diluted between two charged termini in tryptic peptides, sensitivity is often enhanced.
Features:
Tryp-N puts the basic centers of peptides (the amino terminus and side chains of K/R) in one place to give:
- Enhanced Sensitivity: Boost sensitivity by simply switching enzymes! Get significantly lower LODs in MRM experiments compared to trypsin, which splits ion current with each transition. For optimized catalase peptide MRMs, Tryp-N yields a median sensitivity increase of 4.5x and an average of 35x.
- High Compatibility: Works efficiently across a wide pH range (5-12) and in the presence of various buffer systems and datergents, useful for different sample types and experimental conditions.
- Excellent Specificity: Achieves over 95% specificity for N-terminal cleavage at lysine and arginine, producing b-ion rich spectrum. This enhances PTM localization and de novo sequencing clarity. Using trypsin and Tryp-N together is akin to 3- and 5-nucleic acid sequencing!
- Exceptional Efficiency: Protein digestion can be achieved in 30 minutes or less, siginificantly speeding up your workflow without compromising on quality.
- Quick & Easy: Simplifies the protocol with rapid digestion at optimal temperatures, reducing preparation time and enabling faster progression to analytical stages.
- Thermal Stability: Perform digestions at elevated temperatures up to 65°C, denaturating even stubborn proteins while inactivating unwanted proteases.
- Metalloprotease Controllability: Easily turn enzymatic activity on or off with metals and EDTA to precisely and reproducibility control the degree of digestion.
The speed, specificity, ease of use and overall effectiveness of Tryp-N protease, combined with its ability to provide more interpretable spectra make it a valuable component of the proteomics toolkit.
¹Wilson JP, Ipsaro JJ, Del Giudice SN, Turna NS, Gauss CM, Dusenbury KH, Marquart K, Rivera KD, Pappin DJ. Tryp-N: A thermostable protease for the production of N-terminal argininyl and lysinyl peptides. Journal of Proteome Research. 2020 Mar 6;19(4):1459-69.
For our full offer of ProtiFi’s Tryp-N products, visit our catalog.
For more information on ProtiFi, click here.