Long-range cooperative interactions modulate dimerization in SARS 3CL pro

Jennifer Barrila, Usman Bacha, Ernesto Freire

Research output: Contribution to journalArticle

28 Citations (Scopus)

Abstract

Severe acute respiratory syndrome (SARS) is an infectious disease caused by the human coronavirus, SARS-CoV. The main viral protease, SARS 3CL pro, is a validated target for the development of antiviral therapies. Since the enzyme is a homodimer and the individual monomers are inactive, two approaches are being used to develop inhibitors: enzyme activity inhibitors that target the active site and dimerization inhibitors. Dimerization inhibitors are usually targeted to the dimerization interface and need to compete with the attractive forces between subunits to be effective. In this paper, we show that the dimerization of SARS 3CLpro is also under allosteric control and that additional and energetically more favorable target sites away from the dimerization interface may also lead to subunit dissociation. We previously identified a cluster of conserved serine residues (Ser139, Ser144, and Ser147) located adjacent to the active site of 3CL pro that could effectively be targeted to inactivate the protease [Bacha, U et al. (2004) Biochemistry 43, 4906-4912]. Mutation of any of these serine residues to alanine had a debilitating effect on the catalytic activity of 3CLpro. In particular, the mutation of Ser147, which does not make any contact with the opposing subunit and is located approximately 9 A away from the dimer interface, totally inhibited dimerization and resulted in a complete loss of enzymatic activity. The finding that residues away from the dimer interface are able to control dimerization defines alternative targets for the design of dimerization inhibitors.

Original languageEnglish (US)
Pages (from-to)14908-14916
Number of pages9
JournalBiochemistry
Volume45
Issue number50
DOIs
StatePublished - Dec 19 2006
Externally publishedYes

Fingerprint

Severe Acute Respiratory Syndrome
Dimerization
Dimers
Serine
Catalytic Domain
Peptide Hydrolases
Coronavirus
Mutation
Biochemistry
Enzyme activity
Enzyme Inhibitors
Alanine
Antiviral Agents
Communicable Diseases
Catalyst activity
Monomers
Enzymes

ASJC Scopus subject areas

  • Biochemistry

Cite this

Long-range cooperative interactions modulate dimerization in SARS 3CL pro . / Barrila, Jennifer; Bacha, Usman; Freire, Ernesto.

In: Biochemistry, Vol. 45, No. 50, 19.12.2006, p. 14908-14916.

Research output: Contribution to journalArticle

Barrila, Jennifer ; Bacha, Usman ; Freire, Ernesto. / Long-range cooperative interactions modulate dimerization in SARS 3CL pro In: Biochemistry. 2006 ; Vol. 45, No. 50. pp. 14908-14916.
@article{0860545cf3674549913ae50ec8805363,
title = "Long-range cooperative interactions modulate dimerization in SARS 3CL pro",
abstract = "Severe acute respiratory syndrome (SARS) is an infectious disease caused by the human coronavirus, SARS-CoV. The main viral protease, SARS 3CL pro, is a validated target for the development of antiviral therapies. Since the enzyme is a homodimer and the individual monomers are inactive, two approaches are being used to develop inhibitors: enzyme activity inhibitors that target the active site and dimerization inhibitors. Dimerization inhibitors are usually targeted to the dimerization interface and need to compete with the attractive forces between subunits to be effective. In this paper, we show that the dimerization of SARS 3CLpro is also under allosteric control and that additional and energetically more favorable target sites away from the dimerization interface may also lead to subunit dissociation. We previously identified a cluster of conserved serine residues (Ser139, Ser144, and Ser147) located adjacent to the active site of 3CL pro that could effectively be targeted to inactivate the protease [Bacha, U et al. (2004) Biochemistry 43, 4906-4912]. Mutation of any of these serine residues to alanine had a debilitating effect on the catalytic activity of 3CLpro. In particular, the mutation of Ser147, which does not make any contact with the opposing subunit and is located approximately 9 A away from the dimer interface, totally inhibited dimerization and resulted in a complete loss of enzymatic activity. The finding that residues away from the dimer interface are able to control dimerization defines alternative targets for the design of dimerization inhibitors.",
author = "Jennifer Barrila and Usman Bacha and Ernesto Freire",
year = "2006",
month = "12",
day = "19",
doi = "10.1021/bi0616302",
language = "English (US)",
volume = "45",
pages = "14908--14916",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "50",

}

TY - JOUR

T1 - Long-range cooperative interactions modulate dimerization in SARS 3CL pro

AU - Barrila, Jennifer

AU - Bacha, Usman

AU - Freire, Ernesto

PY - 2006/12/19

Y1 - 2006/12/19

N2 - Severe acute respiratory syndrome (SARS) is an infectious disease caused by the human coronavirus, SARS-CoV. The main viral protease, SARS 3CL pro, is a validated target for the development of antiviral therapies. Since the enzyme is a homodimer and the individual monomers are inactive, two approaches are being used to develop inhibitors: enzyme activity inhibitors that target the active site and dimerization inhibitors. Dimerization inhibitors are usually targeted to the dimerization interface and need to compete with the attractive forces between subunits to be effective. In this paper, we show that the dimerization of SARS 3CLpro is also under allosteric control and that additional and energetically more favorable target sites away from the dimerization interface may also lead to subunit dissociation. We previously identified a cluster of conserved serine residues (Ser139, Ser144, and Ser147) located adjacent to the active site of 3CL pro that could effectively be targeted to inactivate the protease [Bacha, U et al. (2004) Biochemistry 43, 4906-4912]. Mutation of any of these serine residues to alanine had a debilitating effect on the catalytic activity of 3CLpro. In particular, the mutation of Ser147, which does not make any contact with the opposing subunit and is located approximately 9 A away from the dimer interface, totally inhibited dimerization and resulted in a complete loss of enzymatic activity. The finding that residues away from the dimer interface are able to control dimerization defines alternative targets for the design of dimerization inhibitors.

AB - Severe acute respiratory syndrome (SARS) is an infectious disease caused by the human coronavirus, SARS-CoV. The main viral protease, SARS 3CL pro, is a validated target for the development of antiviral therapies. Since the enzyme is a homodimer and the individual monomers are inactive, two approaches are being used to develop inhibitors: enzyme activity inhibitors that target the active site and dimerization inhibitors. Dimerization inhibitors are usually targeted to the dimerization interface and need to compete with the attractive forces between subunits to be effective. In this paper, we show that the dimerization of SARS 3CLpro is also under allosteric control and that additional and energetically more favorable target sites away from the dimerization interface may also lead to subunit dissociation. We previously identified a cluster of conserved serine residues (Ser139, Ser144, and Ser147) located adjacent to the active site of 3CL pro that could effectively be targeted to inactivate the protease [Bacha, U et al. (2004) Biochemistry 43, 4906-4912]. Mutation of any of these serine residues to alanine had a debilitating effect on the catalytic activity of 3CLpro. In particular, the mutation of Ser147, which does not make any contact with the opposing subunit and is located approximately 9 A away from the dimer interface, totally inhibited dimerization and resulted in a complete loss of enzymatic activity. The finding that residues away from the dimer interface are able to control dimerization defines alternative targets for the design of dimerization inhibitors.

UR - http://www.scopus.com/inward/record.url?scp=33845585220&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33845585220&partnerID=8YFLogxK

U2 - 10.1021/bi0616302

DO - 10.1021/bi0616302

M3 - Article

VL - 45

SP - 14908

EP - 14916

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 50

ER -