Sequence-specific cleavage of protein fusions using a recombinant Neisseria type 2 IgA protease

Sequence-specific enzymatic cleavage of protein fusions is an important application in recombinant protein technology. We have used the Neisseria type 2 IgA protease (EC 3.4.24.13), produced and secreted by Escherichia coli host cells, for efficiently processing polypeptides at authentic or engineered target sites. In different substrates, the microbial protease specifically cleaves the peptide bond distal to the second Pro residue of the sequence Yaa-Pro-/-Xaa-Pro, where Yaa stands for Pro (or rarely for Pro in combination with Ala, Gly or Thr) and Xaa stands for Thr, Ser or Ala. Highly specific proteolysis has been obtained not only with soluble and purified protein fusions but also with insoluble aggregates derived from cytoplasmic inclusion bodies. The sequence-specificity and simple production of the recombinant IgA protease make it a versatile tool for the in vitro processing of recombinant proteins.