Immobilized protease-assisted synthesis of engineered cysteine-knot microproteins

Abstract

(Chemical Equation Presented) Engineered cyclotides. The exceptionally potent trypsin inhibitor macrocyclic cysteine-knot microproteins (or "cyclotides") MCoTI-I and MCoTI-II and analogues were synthesised by one-pot protease-mediated backbone ligation/ affinity purification and/or thia-zip native chemical ligation cascade cyclization. One of these MCoTI analogues (MCoTI-II[K10F]) is the first example of an engineered MCoTI cyclotide in which specificity is redirected towards an alternative enzyme target (chymotrypsin), whilst retaining high affinity. © 2007 Wiley-VCH Verlag GmbH & Co. KGaA.