Abstract
We identified small-molecule dimer disruptors that inhibit an essential dimeric protease of human Kaposi's sarcoma-associated herpesvirus (KSHV) by screening an alpha-helical mimetic library. Next, we synthesized a second generation of low-micromolar inhibitors with improved potency and solubility. Complementary methods including size exclusion chromatography and 1H-13C HSQC titration using selectively labeled 13C-Met samples revealed that monomeric protease is enriched in the presence of inhibitor. 1H-15N HSQC titration studies mapped the inhibitor binding site to the dimer interface, and mutagenesis studies targeting this region were consistent with a mechanism where inhibitor binding prevents dimerization through the conformational selection of a dynamic intermediate. These results validate the interface of herpesvirus proteases and other similar oligomeric interactions as suitable targets for the development of small-molecule inhibitors.
MeSH terms
-
Binding Sites
-
Herpesvirus 8, Human / drug effects*
-
Herpesvirus 8, Human / enzymology*
-
Herpesvirus 8, Human / genetics
-
Humans
-
Models, Molecular
-
Point Mutation
-
Protease Inhibitors / chemical synthesis
-
Protease Inhibitors / chemistry
-
Protease Inhibitors / pharmacology*
-
Protein Conformation
-
Protein Multimerization / drug effects*
-
Small Molecule Libraries / chemical synthesis
-
Small Molecule Libraries / chemistry
-
Small Molecule Libraries / pharmacology*
-
Substrate Specificity
Substances
-
Protease Inhibitors
-
Small Molecule Libraries
Associated data
-
PubChem-Substance/81055042
-
PubChem-Substance/81055043
-
PubChem-Substance/81067414
-
PubChem-Substance/81067415
-
PubChem-Substance/81067416
-
PubChem-Substance/81067417
-
PubChem-Substance/81067418
-
PubChem-Substance/81067419
-
PubChem-Substance/81067420
-
PubChem-Substance/81067421
-
PubChem-Substance/81067422
-
PubChem-Substance/81067423
-
PubChem-Substance/81067424
-
PubChem-Substance/81067425
-
PubChem-Substance/81067426
-
PubChem-Substance/81067427
-
PubChem-Substance/81067428
-
PubChem-Substance/81067429
-
PubChem-Substance/81067430
-
PubChem-Substance/81067431
-
PubChem-Substance/81067432
-
PubChem-Substance/81067433
-
PubChem-Substance/81067434
-
PubChem-Substance/81067435
-
PubChem-Substance/81067436
-
PubChem-Substance/81067437