Comparison of the substrate specificity of two potyvirus proteases

J. Tőzsér, Joseph E. Tropea, Scott Cherry, P. Bagossi, Terry D. Copeland, Alexander Wlodawer, David S. Waugh

Research output: Contribution to journalArticle

34 Citations (Scopus)

Abstract

The substrate specificity of the nuclear inclusion protein a (NIa) proteolytic enzymes from two potyviruses, the tobacco etch virus (TEV) and tobacco vein mottling virus (TVMV), was compared using oligopeptide substrates. Mutations were introduced into TEV protease in an effort to identify key determinants of substrate specificity. The specificity of the mutant enzymes was assessed by using peptides with complementary substitutions. The crystal structure of TEV protease and a homology model of TVMV protease were used to interpret the kinetic data. A comparison of the two structures and the experimental data suggested that the differences in the specificity of the two enzymes may be mainly due to the variation in their S4 and S3 binding subsites. Two key residues predicted to be important for these differences were replaced in TEV protease with the corresponding residues of TVMV protease. Kinetic analyses of the mutants confirmed that these residues play a role in the specificity of the two enzymes. Additional residues in the substrate-binding subsites of TEV protease were also mutated in an effort to alter the specificity of the enzyme.

Original languageEnglish
Pages (from-to)514-523
Number of pages10
JournalFEBS Journal
Volume272
Issue number2
DOIs
Publication statusPublished - Jan 2005

Fingerprint

Potyvirus
Tobacco
Substrate Specificity
Viruses
Peptide Hydrolases
Veins
Substrates
Enzymes
Intranuclear Inclusion Bodies
Oligopeptides
Kinetics
Nuclear Proteins
Substitution reactions
Crystal structure
Peptides
Mutation
TEV protease
Proteins

Keywords

  • Nuclear inclusion protease
  • Potyvirus protease
  • Substrate specificity
  • Tobacco etch virus protease
  • Tobacco vein mottling virus protease

ASJC Scopus subject areas

  • Biochemistry

Cite this

Tőzsér, J., Tropea, J. E., Cherry, S., Bagossi, P., Copeland, T. D., Wlodawer, A., & Waugh, D. S. (2005). Comparison of the substrate specificity of two potyvirus proteases. FEBS Journal, 272(2), 514-523. https://doi.org/10.1111/j.1742-4658.2004.04493.x

Comparison of the substrate specificity of two potyvirus proteases. / Tőzsér, J.; Tropea, Joseph E.; Cherry, Scott; Bagossi, P.; Copeland, Terry D.; Wlodawer, Alexander; Waugh, David S.

In: FEBS Journal, Vol. 272, No. 2, 01.2005, p. 514-523.

Research output: Contribution to journalArticle

Tőzsér, J, Tropea, JE, Cherry, S, Bagossi, P, Copeland, TD, Wlodawer, A & Waugh, DS 2005, 'Comparison of the substrate specificity of two potyvirus proteases', FEBS Journal, vol. 272, no. 2, pp. 514-523. https://doi.org/10.1111/j.1742-4658.2004.04493.x
Tőzsér, J. ; Tropea, Joseph E. ; Cherry, Scott ; Bagossi, P. ; Copeland, Terry D. ; Wlodawer, Alexander ; Waugh, David S. / Comparison of the substrate specificity of two potyvirus proteases. In: FEBS Journal. 2005 ; Vol. 272, No. 2. pp. 514-523.
@article{0876a0586e774333b7ec6e1687702ae5,
title = "Comparison of the substrate specificity of two potyvirus proteases",
abstract = "The substrate specificity of the nuclear inclusion protein a (NIa) proteolytic enzymes from two potyviruses, the tobacco etch virus (TEV) and tobacco vein mottling virus (TVMV), was compared using oligopeptide substrates. Mutations were introduced into TEV protease in an effort to identify key determinants of substrate specificity. The specificity of the mutant enzymes was assessed by using peptides with complementary substitutions. The crystal structure of TEV protease and a homology model of TVMV protease were used to interpret the kinetic data. A comparison of the two structures and the experimental data suggested that the differences in the specificity of the two enzymes may be mainly due to the variation in their S4 and S3 binding subsites. Two key residues predicted to be important for these differences were replaced in TEV protease with the corresponding residues of TVMV protease. Kinetic analyses of the mutants confirmed that these residues play a role in the specificity of the two enzymes. Additional residues in the substrate-binding subsites of TEV protease were also mutated in an effort to alter the specificity of the enzyme.",
keywords = "Nuclear inclusion protease, Potyvirus protease, Substrate specificity, Tobacco etch virus protease, Tobacco vein mottling virus protease",
author = "J. Tőzs{\'e}r and Tropea, {Joseph E.} and Scott Cherry and P. Bagossi and Copeland, {Terry D.} and Alexander Wlodawer and Waugh, {David S.}",
year = "2005",
month = "1",
doi = "10.1111/j.1742-4658.2004.04493.x",
language = "English",
volume = "272",
pages = "514--523",
journal = "FEBS Journal",
issn = "1742-464X",
publisher = "Wiley-Blackwell",
number = "2",

}

TY - JOUR

T1 - Comparison of the substrate specificity of two potyvirus proteases

AU - Tőzsér, J.

AU - Tropea, Joseph E.

AU - Cherry, Scott

AU - Bagossi, P.

AU - Copeland, Terry D.

AU - Wlodawer, Alexander

AU - Waugh, David S.

PY - 2005/1

Y1 - 2005/1

N2 - The substrate specificity of the nuclear inclusion protein a (NIa) proteolytic enzymes from two potyviruses, the tobacco etch virus (TEV) and tobacco vein mottling virus (TVMV), was compared using oligopeptide substrates. Mutations were introduced into TEV protease in an effort to identify key determinants of substrate specificity. The specificity of the mutant enzymes was assessed by using peptides with complementary substitutions. The crystal structure of TEV protease and a homology model of TVMV protease were used to interpret the kinetic data. A comparison of the two structures and the experimental data suggested that the differences in the specificity of the two enzymes may be mainly due to the variation in their S4 and S3 binding subsites. Two key residues predicted to be important for these differences were replaced in TEV protease with the corresponding residues of TVMV protease. Kinetic analyses of the mutants confirmed that these residues play a role in the specificity of the two enzymes. Additional residues in the substrate-binding subsites of TEV protease were also mutated in an effort to alter the specificity of the enzyme.

AB - The substrate specificity of the nuclear inclusion protein a (NIa) proteolytic enzymes from two potyviruses, the tobacco etch virus (TEV) and tobacco vein mottling virus (TVMV), was compared using oligopeptide substrates. Mutations were introduced into TEV protease in an effort to identify key determinants of substrate specificity. The specificity of the mutant enzymes was assessed by using peptides with complementary substitutions. The crystal structure of TEV protease and a homology model of TVMV protease were used to interpret the kinetic data. A comparison of the two structures and the experimental data suggested that the differences in the specificity of the two enzymes may be mainly due to the variation in their S4 and S3 binding subsites. Two key residues predicted to be important for these differences were replaced in TEV protease with the corresponding residues of TVMV protease. Kinetic analyses of the mutants confirmed that these residues play a role in the specificity of the two enzymes. Additional residues in the substrate-binding subsites of TEV protease were also mutated in an effort to alter the specificity of the enzyme.

KW - Nuclear inclusion protease

KW - Potyvirus protease

KW - Substrate specificity

KW - Tobacco etch virus protease

KW - Tobacco vein mottling virus protease

UR - http://www.scopus.com/inward/record.url?scp=12544254189&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=12544254189&partnerID=8YFLogxK

U2 - 10.1111/j.1742-4658.2004.04493.x

DO - 10.1111/j.1742-4658.2004.04493.x

M3 - Article

VL - 272

SP - 514

EP - 523

JO - FEBS Journal

JF - FEBS Journal

SN - 1742-464X

IS - 2

ER -