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Tryp-N: A thermostable protease for the production of N-terminal argininyl and lysinyl peptides

ABSTRACT: Identification and characterization of Tryp-N, A thermostable protease for the production of N-terminal argininyl and lysinyl peptides

INSTRUMENT(S): LTQ Orbitrap XL

ORGANISM(S): Bos Taurus (bovine) Escherichia Coli Gallus Gallus (chicken) Oryctolagus Cuniculus (rabbit) Saccharomyces Cerevisiae (baker's Yeast) Chaetomium Thermophilum (strain Dsm 1495 / Cbs 144.50 / Imi 039719)

SUBMITTER: Jonathan Ipsaro

LAB HEAD: Darryl J. Pappin

PROVIDER: PXD017024 | Pride | 2020-04-22

REPOSITORIES: pride

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			Action	DRS
	50C-AA-0min_ND.RAW	Raw
	50C-AA-0min_ND.dat	Other
	50C-AA-0min_RG.RAW	Raw
	50C-AA-0min_RG.dat	Other
	50C-AA-16hr_ND.RAW	Raw

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Publications

Tryp-N: A Thermostable Protease for the Production of N-terminal Argininyl and Lysinyl Peptides.

Wilson John P JP Ipsaro Jonathan J JJ Del Giudice Samantha N SN Turna Nikita Saha NS Gauss Carla M CM Dusenbury Katharine H KH Marquart Krisann K Rivera Keith D KD Pappin Darryl J DJ

Journal of proteome research 20200320 4

Bottom-up proteomics is a mainstay in protein identification and analysis. These studies typically employ proteolytic treatment of biological samples to generate suitably sized peptides for tandem mass spectrometric (MS) analysis. In MS, fragmentation of peptides is largely driven by charge localization. Consequently, peptides with basic centers exclusively on their N-termini produce mainly b-ions. Thus, it was long ago realized that proteases that yield such peptides would be valuable proteomic ...[more]

PMID: 32141294

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Tryp-N: A thermostable protease for the production of N-terminal argininyl and lysinyl peptides

Project description:Identification and characterization of Tryp-N, A thermostable protease for the production of N-terminal argininyl and lysinyl peptides

2020-04-22 | PXD017048 | Pride

Tryp-N: A thermostable protease for the production of N-terminal argininyl and lysinyl peptides

Project description:Identification and characterization of Tryp-N, A thermostable protease for the production of N-terminal argininyl and lysinyl peptides

2020-04-22 | PXD017021 | Pride

Tryp-N: A thermostable protease for the production of N-terminal argininyl and lysinyl peptides

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2020-04-22 | PXD017023 | Pride

Tryp-N: A thermostable protease for the production of N-terminal argininyl and lysinyl peptides

Project description:Identification and characterization of Tryp-N, A thermostable protease for the production of N-terminal argininyl and lysinyl peptides

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Tryp-N: A Thermostable Protease for the Production of N-terminal Argininyl and Lysinyl Peptides.

Project description:Bottom-up proteomics is a mainstay in protein identification and analysis. These studies typically employ proteolytic treatment of biological samples to generate suitably sized peptides for tandem mass spectrometric (MS) analysis. In MS, fragmentation of peptides is largely driven by charge localization. Consequently, peptides with basic centers exclusively on their N-termini produce mainly b-ions. Thus, it was long ago realized that proteases that yield such peptides would be valuable proteomic tools for achieving simplified peptide fragmentation patterns and peptide assignment. Work by several groups has identified such proteases, however, structural analysis of these suggested that enzymatic optimization was possible. We therefore endeavored to find enzymes that could provide enhanced activity and versatility while maintaining specificity. Using these previously described proteases as informatic search templates, we discovered and then characterized a thermophilic metalloprotease with N-terminal specificity for arginine and lysine. This enzyme, dubbed Tryp-N, affords many advantages including improved thermostability, solvent and detergent tolerance, and rapid digestion time.

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