Substrate Specificities of Porcine and Bovine Enteropeptidases toward the Peptide Val-(Asp)₄-Lys-Ile-Val-Gly and Its Analogs

An overview of enzymatic reagents for the removal of affinity tags

“…Only proline and tryptophan were not well tolerated in the P1 0 position. Another study examining the importance of the P1-P5 positions concluded that the P1 lysine was the most important specificity determinant, followed by the aspartate residues in the P2, P3, P5 and P4 positions, respectively, with the latter position contributing very little to specificity [27]. Interestingly these investigators found that the sequence DDDDR was cleaved more efficiently than the canonical DDDDK.…”

Membrane-Anchored Serine Proteases in Health and Disease

“…Within this recognition sequence, a Lys or an Arg residue at P 1 and Lys residues at P 2 and P 3 appear to be most important for efficient cleavage. 87 The structural determinants for enteropeptidase substrate specificity have been localized in its protease domain. There is a group of four conserved basic residues, R/KRRK at positions 96-99, which were suspected to interact with the acidic P 2 -P 5 residues in the trypsinogen activation site.…”

Importance of a single disulfide bond for the PsbO protein of photosystem II: protein structure stability and soluble overexpression in Escherichia coli

“…Generally enterokinase cleaves proteins specifically, but in a number of studies non-specifical cleavage of recombinant proteins by enterokinase in positions different from the canonical (Asp) 4 Lys; sequence has been reported (Uegaki et al 1996;Safi et al 2001;Agnihotri et al 2001). Recent work has demonstrated that enterokinase can recognize and hydrolyze substrates containing less than four (from one to three) negatively charged residues (Asp/Glu) upstream of Lys (Likhareva et al 2003;Kim et al 2008). The general formula of such substrates is-Asp m (Glu) m -X n -Lys;-, where X-Asp, Glu or any other amino acid residue, m = 0-1, n = 0-3.…”