Cleavage Site
Definition:
This sequence is positioned between the fusion tag/protein and the target protein where it is recognized by proteases or other cleaving agents. Although the fusion tag/protein is often important for protein production and/or purification, it may interfere with: Addition of proteases or cleaving agents which specifically recognize the cleaving site, allow for easy and efficient release of the target protein from the fusion tag/protein.
 
Frequently Used Proteases and Other Cleaving Agents
Related DBs & Software
Some commercially available plasmids with cleavage sites

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Relevant DBs & Software Related to Cleavage Sites & Cleaving Enzymes

When choosing vectors for expression, containing appropriate cleavage sites, it is important to ensure that the chosen cleavage site is unique to the sequence. In order to ensure that it is not included within the target protein sequence, the Peptide Cutter is recommended. This tool allows one to enter a protein sequence and receive a list of potential cleavage sites (note that not all proteases are recognized by this program).

Brenda - presents enzyme functional data, and is maintained and developed at the institute of Biochemistry at the University of Cologne.
MEROPS - the protease database
EPD - the E-coli proteases database (Ehrmann's lab in Cardiff University).
Peptide Cutter - predicts potential protease and cleavage sites and sites cleaved by chemicals in a given protein sequence.

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Frequently Used Proteases and Other Cleaving Agents
(For additional information about other cleaving agents see: Peptide cutter introduction )
Cleaving Agent 
(Linked to the 
BRENDA DATABASE* )
 Recognized Sequence
(Purple line represents site of cleavage) 
Related Procedures
(See also: Dr. Lebendiker's site )
Related Literature 
Enterokinase/Enteropeptidase
3.4.21.9
Asp Asp Asp Asp Lys |
  1. NOVAGEN Enterokinase cleavage capture kit
  2. ROCHE Enterokinase (pdf)
 Suppliers of: Enterokinase
(Using BioSupplyNet)
Factor Xa
3.4.21.6
Ile Glu/Asp Gly Arg |
NOVAGEN-Xa kit (pdf)
  1. A critical review of the methods for cleavage of fusion proteins with thrombin and factor Xa
  2. Suppliers of: Factor Xa (Using BioSupplyNet)

Sumo Protease
3.4.22.68

Gly-Gly-|-Ala-Thr-Tyr
at the C-terminal end of SUMO

The enzyme recognizes the tertiary structure of SUMO

 
TEV (Tobacco Etch Virus) protease 
[= nuclear inclusion A endopeptidase]
3.4.22.44
Glu Asn Leu Tyr Phe Gln |Gly
  1. INVITROGEN- LIFE TECHNOLOGIES 
  2. TEV-Protease (pdf) 
  1. General Information (pdf)  
  2. Large scale purification of a stable form of TEV (abstract)  
Thrombin
3.4.21.5 
Leu Val Pro Arg |Gly Ser
  1. Thrombin Cleavage of a GST fusion protein
  2. NOVAGEN Thrombin kit (PDF)
  1. A critical review of the methods for cleavage of fusion proteins with thrombin and factor Xa.
  2. Suppliers of: Thrombin (Using BioSupplyNet)
Signal Peptidase I
3.4.21.89
 Ala X Ala
  
Additional information 
Detergent: DTT in Chitin binding kit (pdf)
 
  
 
Detergent: Cyanogen Bromide
X Met | X
    
* The BRENDA DATABASE offers valuable information relating to various molecular and functional paramters characterizing enzymes.

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