Conformational changes in human hepatitis C virus NS3 protease upon binding of product-based inhibitors

Elisabetta Bianchi, Stefania Orrù, Fabrizio Dal Piaz, Raffaele Ingenito, Annarita Casbarra, Gabriella Biasiol, Uwe Koch, Piero Pucci, Antonello Pessi

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

One of the most promising approaches to anti-hepatitis C virus drug discovery is the development of inhibitors of the virally encoded protease NS3. This chymotrypsin-like serine protease as essential for the maturation of the viral polyprotein, and processing requires complex formation between NS3 and its cofactor NS4A. Recently, we reported on the discovery of potent cleavage product-derived inhibitors [Ingallinella et al. (1998) Biochemistry 37, 8906-8914]. Here we study the interaction of these inhibitors with NS3 and the NS3/cofactor complex. Inhibitors bind NS3 according to an induced-fit mechanism. In the absence of cofactor different binding modes are apparent, while in the presence of cofactor all inhibitors show the same binding mode with a small rearrangement in the NS3 structure, as suggested by circular dichroism spectroscopy. These data are consistent with the hypothesis that NS4A complexation induces an NS3 structure that is already (but not entirely) preorganized for substrate binding not only for what concerns the S' site, as already suggested, but also for the S site. Inhibitor binding to the NS3/cofactor complex induces the stabilization of the enzyme structure as highlighted by limited proteolysis experiments. We envisage that this may occur through stabilization of the individual N-terminal and C-terminal domains where the cofactor and inhibitor, respectively, bind and subsequent tightening of the interdomain interaction in the ternary complex.

Original languageEnglish
Pages (from-to)13844-13852
Number of pages9
JournalBiochemistry
Volume38
Issue number42
DOIs
Publication statusPublished - Oct 19 1999

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Chymases
Polyproteins
Serine Proteases
Drug Discovery
Circular Dichroism
Hepacivirus
Biochemistry
Proteolysis
Spectrum Analysis
Peptide Hydrolases
Stabilization
Circular dichroism spectroscopy
Chymotrypsin
Enzymes
Complexation
Viruses
Substrates
Processing
Experiments
hepatitis C virus NS3 protein

ASJC Scopus subject areas

  • Biochemistry

Cite this

Bianchi, E., Orrù, S., Dal Piaz, F., Ingenito, R., Casbarra, A., Biasiol, G., ... Pessi, A. (1999). Conformational changes in human hepatitis C virus NS3 protease upon binding of product-based inhibitors. Biochemistry, 38(42), 13844-13852. https://doi.org/10.1021/bi991220w

Conformational changes in human hepatitis C virus NS3 protease upon binding of product-based inhibitors. / Bianchi, Elisabetta; Orrù, Stefania; Dal Piaz, Fabrizio; Ingenito, Raffaele; Casbarra, Annarita; Biasiol, Gabriella; Koch, Uwe; Pucci, Piero; Pessi, Antonello.

In: Biochemistry, Vol. 38, No. 42, 19.10.1999, p. 13844-13852.

Research output: Contribution to journalArticle

Bianchi, E, Orrù, S, Dal Piaz, F, Ingenito, R, Casbarra, A, Biasiol, G, Koch, U, Pucci, P & Pessi, A 1999, 'Conformational changes in human hepatitis C virus NS3 protease upon binding of product-based inhibitors', Biochemistry, vol. 38, no. 42, pp. 13844-13852. https://doi.org/10.1021/bi991220w
Bianchi, Elisabetta ; Orrù, Stefania ; Dal Piaz, Fabrizio ; Ingenito, Raffaele ; Casbarra, Annarita ; Biasiol, Gabriella ; Koch, Uwe ; Pucci, Piero ; Pessi, Antonello. / Conformational changes in human hepatitis C virus NS3 protease upon binding of product-based inhibitors. In: Biochemistry. 1999 ; Vol. 38, No. 42. pp. 13844-13852.
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