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Frequently used protein cleavage reagents |
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| Enzymes |
Preferred cleavage site |
Approximate frequency |
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| Ancrod |
Arg-X, Arg-Gly |
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| Bromelain |
C-terminal to Lys, Ala and Tyr |
|
| Chymotrypsin |
C-terminal to hydrophobic residues, e.g., Phe, Tyr, Trp. Less sensitive with Leu, Met, Ala |
|
| Clostripain |
C-terminal to Arg residues |
20 |
| Collagenase |
N-terminal to Gly (X-Gly) in Pro-X-Gly-Pro |
|
| Elastase |
C-terminal to amino acids with small hydrophobic side chains |
|
| Endoproteinase Arg-C |
C-terminal to Arg residues |
20 |
| Endoproteinase Asp-N |
N-terminal to Asp and Cys |
10 |
| Endoproteinase Glu-C |
C-terminal to Asp and Glu |
10 |
| Endoproteinase Lys-C |
C-terminal to Lys |
20 |
| Factor Xa |
C-terminal to Arg in Gly-Arg-X |
|
| Ficin |
uncharged or aromatic amino acids |
|
| Follipsin |
Arg-X |
20 |
| Kallikrein |
C-terminal to Arg in (Phe-Arg-X or Leu-Arg-X) |
|
| Pepsin |
Broad specificity; preference for cleavage C-terminal to Phe, Leu, and Glu |
7 |
| Thermolysin |
N-terminal to amino acids with bulky hydrophobic side chains, e.g., Ile, Leu, Val, and Phe 5 |
|
| Thrombin |
C-terminal to Arg |
|
| Trypsin |
C-terminal to Lys and Arg |
10 |
| V8 protease |
C-terminal to Glu, less active with Asp |
|
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| Chemistry |
Preferred cleavage site |
|
|
|
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| Cyanogen bromide |
Trp, (Met) |
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| Formic acid |
Asp – Pro |
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| HCl |
Asp-X, X-Asp |
|
| Hydroxylamine (alkaline pH) |
Asn – Gly |
|
| N-bromosuccinimide (NBS) or N- chlorosuccinimide |
Trp |
|
| 2-Nitro-5-thiocyanobenzoate (NTCB) |
Cys |
|
Soloviev et al. Journal of Nanobiotechnology 2003 1:4 doi:10.1186/1477-3155-1-4 |
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