Engineered staphylococcal protein A's IgG-binding domain with cathepsin L inhibitory activity.
Inhibitory peptide of papain-like cysteine proteases, affinity selected from a random disulfide constrained phage-displayed peptide library, was grafted to staphylococcal protein A's B domain. Scaffold protein was additionally modified in order to allow solvent exposed display of peptide loop. Correct folding of fusion proteins was confirmed by CD-spectroscopy and by the ability to bind the Fc-region of rabbit IgG, a characteristic of parent domain. The recombinant constructs inhibited cathepsin L with inhibitory constants in the low-micromolar range.
Bratkovic
Tomaz
T
Faculty of Pharmacy, Department of Pharmaceutical Biology, University of Ljubljana, Askerceva 7, SI-1000 Ljubljana, Slovenia. tomaz.bratkovic@ffa.uni-lj.si
Berlec
Ales
A
Popovic
Tatjana
T
Lunder
Mojca
M
Kreft
Samo
S
Urleb
Uros
U
Strukelj
Borut
B
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D056668:Cathepsin L; D002403:Cathepsins / Q000037:antagonists & inhibitors* / Q000737:chemistry*; D003546:Cysteine Endopeptidases / Q000737:chemistry*; D004220:Disulfides / Q000737:chemistry; D007074:Immunoglobulin G / Q000737:chemistry*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D019151:Peptide Library; D011485:Protein Binding; D017510:Protein Folding; D017434:Protein Structure, Tertiary; D011817:Rabbits; D013205:Staphylococcal Protein A / Q000737:chemistry*
D004220:Disulfides; D007074:Immunoglobulin G; D019151:Peptide Library; D013205:Staphylococcal Protein A; D002403:Cathepsins; D003546:Cysteine Endopeptidases; D056668:Cathepsin L