Influence of the amino acid residue downstream of (Asp)4Lys on enterokinase cleavage of a fusion protein.
We have studied the cleavage efficiency of the protease enterokinase (EK) using the novel vector pESP4. pESP4 is a yeast expression vector equipped with ligation-independent cloning sites, a GST purification tag, and a FLAG epitope tag. EK is used to cleave the FLAG and GST tags leaving the protein of interest without any extraneously added amino acids. We have found that EK is relatively permissive of the amino acid residue downstream of the recognition sequence (the P'1 position). This makes EK an ideal choice to use as a protease to cleave any protein of interest cloned within the pESP4 yeast expression vector.
Hosfield
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Stratagene Cloning Systems, Inc., La Jolla, California 92037, USA. tanya_hosfield@stratagene.com
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D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D002147:Calmodulin / Q000235:genetics / Q000378:metabolism*; D003001:Cloning, Molecular; D004765:Enteropeptidase / Q000378:metabolism*; D005822:Genetic Vectors; D005982:Glutathione Transferase / Q000235:genetics / Q000378:metabolism*; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000737:chemistry*; D011487:Protein Conformation; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification / Q000378:metabolism; D012568:Schizosaccharomyces
D002147:Calmodulin; D009842:Oligopeptides; D011993:Recombinant Fusion Proteins; C080396:trypsinogen activation peptide; D005982:Glutathione Transferase; D004765:Enteropeptidase