Engineered cystine knot miniproteins as potent inhibitors of human mast cell tryptase beta.
Here we report the design, chemical and recombinant synthesis, and functional properties of a series of novel inhibitors of human mast cell tryptase beta, a protease of considerable interest as a therapeutic target for the treatment of allergic asthma and inflammatory disorders. These inhibitors are derived from a linear variant of the cyclic cystine knot miniprotein MCoTI-II, originally isolated from the seeds of Momordica cochinchinensis. A synthetic cyclic miniprotein that bears additional positive charge in the loop connecting the N- and C-termini inhibits all monomers of the tryptase beta tetramer with an overall equilibrium dissociation constant K(i) of 1 nM and thus is one of the most potent proteinaceous inhibitors of tryptase beta described to date. These cystine knot miniproteins may therefore become valuable scaffolds for the design of a new generation of tryptase inhibitors.
Sommerhoff
Christian P
CP
Division of Clinical Chemistry and Clinical Biochemistry, Surgical Department, Ludwig-Maximilians-University Munich, Nussbaumstrasse 20, D-80336 Munich, Germany.
Avrutina
Olga
O
Schmoldt
Hans-Ulrich
HU
Gabrijelcic-Geiger
Dusica
D
Diederichsen
Ulf
U
Kolmar
Harald
H
D000595:Amino Acid Sequence; D000818:Animals; D002417:Cattle; D047169:Cyclotides / Q000737:chemistry* / Q000494:pharmacology*; D047168:Cystine Knot Motifs; D006801:Humans; D008958:Models, Molecular; D008969:Molecular Sequence Data; D015202:Protein Engineering; D011993:Recombinant Fusion Proteins / Q000737:chemistry; D017386:Sequence Homology, Amino Acid; D040681:Structural Homology, Protein; D053802:Tryptases / Q000037:antagonists & inhibitors* / Q000737:chemistry
D047169:Cyclotides; C519458:McoEeTI protein; D011993:Recombinant Fusion Proteins; C409923:trypsin inhibitor MCoTI-II; D053802:Tryptases