Endoproteinase Asp-N

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Endopeptidase

Endopeptidases mainly act on peptide bonds within protein polypeptide chains to break down long protein chains into short peptide fragments. The specificity of the cleavage site required by this protease is called the substrate specificity of the protease. The substrate specificity of a protease can be used to obtain peptides with a specific structure in a targeted manner.

Endopeptidase

Endoproteinase Asp-N is an aspartic protease that specifically hydrolyzes the amino terminus of Asp and has promising applications in proteomics analysis. Endoproteinase Asp-N is a zinc metalloendopeptidase. It is highly specific and cleaves peptide bonds on the N-terminal side of aspartic acid residues, and to a lesser extent, glutamic acid (E) residues. It is derived from a mutant strain of Pseudomonas fragi, and has a molecular mass of 27 kDa. In the presence of zinc, it shows optimal activity in a pH range of 4–9 and can remain active under denaturing conditions. Asp-N Protease can be used in combination with other proteases, such as trypsin, for improved protein digestion. Mass spectrometry grade Asp-N protease can be used for peptide matching, protein identification, in-solution and in-gel digestion of proteins, and peptide mass fingerprinting. Visit the enzyme page for more product information.

Source

This Asp-N is a mass spectrometry (MS)-grade zinc metalloproteinase derived from a mutant strain of Pseudomonas fragi and requires a trace amount of zinc for activity. Asp-N can be used alone or in parallel with trypsin or other proteases to produce protein digests for peptide mapping and protein sequencing. Asp-N protease is suitable for either in-solution or in-gel digestion workflows.

Properties

The endoproteinase Asp-N cleaves primarily at amino side of aspartate residues and cysteic acid residues that result from the oxidization of cysteine residues, generating a limited number of peptide fragments. Cleavage can also occur at glutamic residues; however, the rate of cleavage at the glutamyl residues is significantly lower than the rate of cleavage at the aspartic acid residues. AspN can efficiently digest protein in 2-20 hours at 37°C. AspN remains active under denaturing conditions such as 1M urea, 2M guanidine·HCl, 0.1% SDS, 2% CHAPS and 10% acetonitrile with optimal activity in the pH range of 6-8. This lyophilized enzyme has a mass of 27 kDa and is stable for 1 year when stored at -20°C.