Prediction of the binding site on E1 in the assembly of the pyruvate dehydrogenase multienzyme complex of Bacillus stearothermophilus

Hyo Il Jung; Richard N. Perham

doi:10.1016/S0014-5793(03)01245-6

Prediction of the binding site on E1 in the assembly of the pyruvate dehydrogenase multienzyme complex of Bacillus stearothermophilus

Hyo Il Jung, Richard N. Perham

Department of Mechanical Engineering

Research output: Contribution to journal › Article › peer-review

5 Citations (Scopus)

Abstract

The β-subunit (E1β) of the pyruvate decarboxylase (E1, α₂β₂) component of the Bacillus stearothermophilus pyruvate dehydrogenase complex was comparatively modelled based on the crystal structures of the homologous 2-oxoisovalerate decarboxylase of Pseudomonas putida and Homo sapiens. Based on this homology modelling, alanine-scanning mutagenesis studies revealed that the negatively charged side chain of Glu285 and the hydrophobic side chain of Phe324 are of particular importance in the interaction with the peripheral subunit-binding domain of the dihydrolipoyl acetyltransferase component of the complex. These results help to identify the site of interaction on the E1β subunit and are consistent with thermodynamic evidence of a mixture of electrostatic and hydrophobic interactions being involved.

Original language	English
Pages (from-to)	405-410
Number of pages	6
Journal	FEBS Letters
Volume	555
Issue number	2
DOIs	https://doi.org/10.1016/S0014-5793(03)01245-6
Publication status	Published - 2003 Dec 4

Bibliographical note

Funding Information:
This work was supported in part by a research grant (to R.N.P.) from the Biotechnology and Biological Sciences Research Council. We are grateful to the BBSRC and The Wellcome Trust for their support of the core facilities in the Cambridge Centre for Molecular Recognition. We thank the Cambridge Overseas Trust, the Department of Biochemistry and St John’s College, Cambridge for financial support to H.I.J.

All Science Journal Classification (ASJC) codes

Biophysics
Structural Biology
Biochemistry
Molecular Biology
Genetics
Cell Biology

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10.1016/S0014-5793(03)01245-6

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title = "Prediction of the binding site on E1 in the assembly of the pyruvate dehydrogenase multienzyme complex of Bacillus stearothermophilus",

abstract = "The β-subunit (E1β) of the pyruvate decarboxylase (E1, α2β2) component of the Bacillus stearothermophilus pyruvate dehydrogenase complex was comparatively modelled based on the crystal structures of the homologous 2-oxoisovalerate decarboxylase of Pseudomonas putida and Homo sapiens. Based on this homology modelling, alanine-scanning mutagenesis studies revealed that the negatively charged side chain of Glu285 and the hydrophobic side chain of Phe324 are of particular importance in the interaction with the peripheral subunit-binding domain of the dihydrolipoyl acetyltransferase component of the complex. These results help to identify the site of interaction on the E1β subunit and are consistent with thermodynamic evidence of a mixture of electrostatic and hydrophobic interactions being involved.",

author = "Jung, {Hyo Il} and Perham, {Richard N.}",

note = "Funding Information: This work was supported in part by a research grant (to R.N.P.) from the Biotechnology and Biological Sciences Research Council. We are grateful to the BBSRC and The Wellcome Trust for their support of the core facilities in the Cambridge Centre for Molecular Recognition. We thank the Cambridge Overseas Trust, the Department of Biochemistry and St John{\textquoteright}s College, Cambridge for financial support to H.I.J. ",

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AU - Perham, Richard N.

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N2 - The β-subunit (E1β) of the pyruvate decarboxylase (E1, α2β2) component of the Bacillus stearothermophilus pyruvate dehydrogenase complex was comparatively modelled based on the crystal structures of the homologous 2-oxoisovalerate decarboxylase of Pseudomonas putida and Homo sapiens. Based on this homology modelling, alanine-scanning mutagenesis studies revealed that the negatively charged side chain of Glu285 and the hydrophobic side chain of Phe324 are of particular importance in the interaction with the peripheral subunit-binding domain of the dihydrolipoyl acetyltransferase component of the complex. These results help to identify the site of interaction on the E1β subunit and are consistent with thermodynamic evidence of a mixture of electrostatic and hydrophobic interactions being involved.

AB - The β-subunit (E1β) of the pyruvate decarboxylase (E1, α2β2) component of the Bacillus stearothermophilus pyruvate dehydrogenase complex was comparatively modelled based on the crystal structures of the homologous 2-oxoisovalerate decarboxylase of Pseudomonas putida and Homo sapiens. Based on this homology modelling, alanine-scanning mutagenesis studies revealed that the negatively charged side chain of Glu285 and the hydrophobic side chain of Phe324 are of particular importance in the interaction with the peripheral subunit-binding domain of the dihydrolipoyl acetyltransferase component of the complex. These results help to identify the site of interaction on the E1β subunit and are consistent with thermodynamic evidence of a mixture of electrostatic and hydrophobic interactions being involved.

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Prediction of the binding site on E1 in the assembly of the pyruvate dehydrogenase multienzyme complex of Bacillus stearothermophilus

Abstract

Bibliographical note

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