Inhibition of a viral enzyme by a small-molecule dimer disruptor.
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.
Shahian
Tina
T
Graduate Group in Biochemistry and Molecular Biology, University of California, San Francisco, California, USA.
Lee
Gregory M
GM
Lazic
Ana
A
Arnold
Leggy A
LA
Velusamy
Priya
P
Roels
Christina M
CM
Guy
R Kiplin
RK
Craik
Charles S
CS
D001665:Binding Sites; D019288:Herpesvirus 8, Human / Q000187:drug effects* / Q000201:enzymology* / Q000235:genetics; D006801:Humans; D008958:Models, Molecular; D017354:Point Mutation; D011480:Protease Inhibitors / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D011487:Protein Conformation; D055503:Protein Multimerization / Q000187:drug effects*; D054852:Small Molecule Libraries / Q000138:chemical synthesis / Q000737:chemistry / Q000494:pharmacology*; D013379:Substrate Specificity
D011480:Protease Inhibitors; D054852:Small Molecule Libraries