Design of cell-permeable, fluorescent activity-based probes for the lysosomal cysteine protease asparaginyl endopeptidase (AEP)/legumain.
Asparaginyl endopeptidase (AEP), also known as legumain, is a cysteine protease that has been ascribed roles in antigen presentation yet its exact role in human biology remains poorly understood. We report here, the use of a positional scanning combinatorial library of peptide AOMKs containing a P1 aspartic acid to probe the P2, P3, and P4 subsite specificity of endogenous legumain. Using inhibitor specificity profiles of cathepsin B and legumain, we designed fluorescent ABPs that are highly selective, cell-permeable reagents for monitoring legumain activity in complex proteomes.
Sexton
Kelly B
KB
Department of Pathology, Stanford University School of Medicine, 300 Pasteur Dr., Stanford, CA 940305, USA.
Witte
Martin D
MD
Blum
Galia
G
Bogyo
Matthew
M
D002401:Cathepsin B / Q000037:antagonists & inhibitors; D002463:Cell Membrane Permeability / Q000502:physiology; D020650:Combinatorial Chemistry Techniques; D003546:Cysteine Endopeptidases / Q000378:metabolism*; D015195:Drug Design; D005456:Fluorescent Dyes / Q000138:chemical synthesis* / Q000494:pharmacology; D008247:Lysosomes / Q000201:enzymology*; D000099240:Asparaginyl Endopeptidase
D002401:Cathepsin B; D003546:Cysteine Endopeptidases; D005456:Fluorescent Dyes; D000099240:Asparaginyl Endopeptidase