The Potency and Specificity of the Interaction between the IA3 Inhibitor and Its Target Aspartic Proteinase from Saccharomyces cerevisiae

Research output: Contribution to journal › Journal article › Research › peer-review

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The Potency and Specificity of the Interaction between the IA3 Inhibitor and Its Target Aspartic Proteinase from Saccharomyces cerevisiae. / Phylip, Lowri H.; Lees, Wendy E.; Brownsey, Brian G.; Bur, Daniel; Dunn, Ben M.; Winther, Jakob R.; Gustchina, Alla; Li, Mi; Copeland, Terry; Wlodawer, Alexander; Kay, John.

In: Journal of Biological Chemistry, Vol. 276, No. 3, 2001, p. 2023-2030.

Research output: Contribution to journal › Journal article › Research › peer-review

Harvard

Phylip, LH, Lees, WE, Brownsey, BG, Bur, D, Dunn, BM, Winther, JR, Gustchina, A, Li, M, Copeland, T, Wlodawer, A & Kay, J 2001, 'The Potency and Specificity of the Interaction between the IA3 Inhibitor and Its Target Aspartic Proteinase from Saccharomyces cerevisiae', Journal of Biological Chemistry, vol. 276, no. 3, pp. 2023-2030. https://doi.org/10.1074/jbc.M008520200

APA

Phylip, L. H., Lees, W. E., Brownsey, B. G., Bur, D., Dunn, B. M., Winther, J. R., Gustchina, A., Li, M., Copeland, T., Wlodawer, A., & Kay, J. (2001). The Potency and Specificity of the Interaction between the IA3 Inhibitor and Its Target Aspartic Proteinase from Saccharomyces cerevisiae. Journal of Biological Chemistry, 276(3), 2023-2030. https://doi.org/10.1074/jbc.M008520200

Vancouver

Phylip LH, Lees WE, Brownsey BG, Bur D, Dunn BM, Winther JR et al. The Potency and Specificity of the Interaction between the IA3 Inhibitor and Its Target Aspartic Proteinase from Saccharomyces cerevisiae. Journal of Biological Chemistry. 2001;276(3):2023-2030. https://doi.org/10.1074/jbc.M008520200

Author

Phylip, Lowri H. ; Lees, Wendy E. ; Brownsey, Brian G. ; Bur, Daniel ; Dunn, Ben M. ; Winther, Jakob R. ; Gustchina, Alla ; Li, Mi ; Copeland, Terry ; Wlodawer, Alexander ; Kay, John. / The Potency and Specificity of the Interaction between the IA3 Inhibitor and Its Target Aspartic Proteinase from Saccharomyces cerevisiae. In: Journal of Biological Chemistry. 2001 ; Vol. 276, No. 3. pp. 2023-2030.

Bibtex

@article{d9bb5f4ca40e4c5ba15c75ddeb165141,
title = "The Potency and Specificity of the Interaction between the IA3 Inhibitor and Its Target Aspartic Proteinase from Saccharomyces cerevisiae",
abstract = "The yeast IA3 polypeptide consists of only 68 residues, and the free inhibitor has little intrinsic secondary structure. IA3 showed subnanomolar potency toward its target, proteinase A from Saccharomyces cerevisiae, and did not inhibit any of a large number of aspartic proteinases with similar sequences/structures from a wide variety of other species. Systematic truncation and mutagenesis of the IA3 polypeptide revealed that the inhibitory activity is located in the N-terminal half of the sequence. Crystal structures of different forms of IA3 complexed with proteinase A showed that residues in the N-terminal half of the IA3 sequence became ordered and formed an almost perfect alpha-helix in the active site of the enzyme. This potent, specific interaction was directed primarily by hydrophobic interactions made by three key features in the inhibitory sequence. Whereas IA3 was cut as a substrate by the nontarget aspartic proteinases, it was not cleaved by proteinase A. The random coil IA3 polypeptide escapes cleavage by being stabilized in a helical conformation upon interaction with the active site of proteinase A. This results, paradoxically, in potent selective inhibition of the target enzyme.",
keywords = "Amino Acid Sequence, Aspartic Acid Endopeptidases, Fungal Proteins, Hydrolysis, Kinetics, Molecular Sequence Data, Peptides, Protease Inhibitors, Protein Conformation, Saccharomyces cerevisiae, Saccharomyces cerevisiae Proteins",
author = "Phylip, {Lowri H.} and Lees, {Wendy E.} and Brownsey, {Brian G.} and Daniel Bur and Dunn, {Ben M.} and Winther, {Jakob R.} and Alla Gustchina and Mi Li and Terry Copeland and Alexander Wlodawer and John Kay",
year = "2001",
doi = "10.1074/jbc.M008520200",
language = "English",
volume = "276",
pages = "2023--2030",
journal = "Journal of Biological Chemistry",
issn = "0021-9258",
publisher = "American Society for Biochemistry and Molecular Biology, Inc.",
number = "3",

}

RIS

TY - JOUR

T1 - The Potency and Specificity of the Interaction between the IA3 Inhibitor and Its Target Aspartic Proteinase from Saccharomyces cerevisiae

AU - Phylip, Lowri H.

AU - Lees, Wendy E.

AU - Brownsey, Brian G.

AU - Bur, Daniel

AU - Dunn, Ben M.

AU - Winther, Jakob R.

AU - Gustchina, Alla

AU - Li, Mi

AU - Copeland, Terry

AU - Wlodawer, Alexander

AU - Kay, John

PY - 2001

Y1 - 2001

N2 - The yeast IA3 polypeptide consists of only 68 residues, and the free inhibitor has little intrinsic secondary structure. IA3 showed subnanomolar potency toward its target, proteinase A from Saccharomyces cerevisiae, and did not inhibit any of a large number of aspartic proteinases with similar sequences/structures from a wide variety of other species. Systematic truncation and mutagenesis of the IA3 polypeptide revealed that the inhibitory activity is located in the N-terminal half of the sequence. Crystal structures of different forms of IA3 complexed with proteinase A showed that residues in the N-terminal half of the IA3 sequence became ordered and formed an almost perfect alpha-helix in the active site of the enzyme. This potent, specific interaction was directed primarily by hydrophobic interactions made by three key features in the inhibitory sequence. Whereas IA3 was cut as a substrate by the nontarget aspartic proteinases, it was not cleaved by proteinase A. The random coil IA3 polypeptide escapes cleavage by being stabilized in a helical conformation upon interaction with the active site of proteinase A. This results, paradoxically, in potent selective inhibition of the target enzyme.

AB - The yeast IA3 polypeptide consists of only 68 residues, and the free inhibitor has little intrinsic secondary structure. IA3 showed subnanomolar potency toward its target, proteinase A from Saccharomyces cerevisiae, and did not inhibit any of a large number of aspartic proteinases with similar sequences/structures from a wide variety of other species. Systematic truncation and mutagenesis of the IA3 polypeptide revealed that the inhibitory activity is located in the N-terminal half of the sequence. Crystal structures of different forms of IA3 complexed with proteinase A showed that residues in the N-terminal half of the IA3 sequence became ordered and formed an almost perfect alpha-helix in the active site of the enzyme. This potent, specific interaction was directed primarily by hydrophobic interactions made by three key features in the inhibitory sequence. Whereas IA3 was cut as a substrate by the nontarget aspartic proteinases, it was not cleaved by proteinase A. The random coil IA3 polypeptide escapes cleavage by being stabilized in a helical conformation upon interaction with the active site of proteinase A. This results, paradoxically, in potent selective inhibition of the target enzyme.

KW - Amino Acid Sequence

KW - Aspartic Acid Endopeptidases

KW - Fungal Proteins

KW - Hydrolysis

KW - Kinetics

KW - Molecular Sequence Data

KW - Peptides

KW - Protease Inhibitors

KW - Protein Conformation

KW - Saccharomyces cerevisiae

KW - Saccharomyces cerevisiae Proteins

U2 - 10.1074/jbc.M008520200

DO - 10.1074/jbc.M008520200

M3 - Journal article

C2 - 11042188

VL - 276

SP - 2023

EP - 2030

JO - Journal of Biological Chemistry

JF - Journal of Biological Chemistry

SN - 0021-9258

IS - 3

ER -

ID: 43973802