Sequential 1H and 15N Nuclear Magnetic Resonance Assignments and Secondary Structure of the Lipoyl Domain of the 2‐Oxoglutarate Dehydrogenase Complex from Azotobacter Vinelandii
- 1 November 1995
- journal article
- Published by Wiley in European Journal of Biochemistry
- Vol. 234 (1) , 148-159
- https://doi.org/10.1111/j.1432-1033.1995.148_c.x
Abstract
A 79‐amino‐acid polypeptide, corresponding to the lipoyl domain of the succinyltransferase component of the 2‐oxoglutarate dehydrogenase multienzyme complex from Azotobacter vinelandii, has been sub‐cloned and produced in Escherichia coli. Complete sequential 1H and 15N resonance assignments for the lipoyl domain have been obtained by using homo‐ and hetero‐nuclear NMR spectroscopy. Two anti‐parallel β‐sheets of four strands each were identified from characteristic NOE connectivities and 3JHNα values. The lipoyl‐lysine residue is found in a type‐I turn connecting two β‐strands. The secondary structure of the lipoyl domain very much resembles the secondary solution structure of the N‐terminal lipoyl domain of the A. vinelandii pyruvate dehydrogenase complex, despite the sequence identity of 25%. A detailed comparison of the NMR‐derived parameters of both lipoyl domains, i.e. chemical shifts, NH‐exchange rates, NOEs, and 3JHNα values suggests a high structural similarity in solution between the two lipoyl domains. Preliminary tertiary‐structure calculations confirm that these lipoyl domains have very similar overall folds. The observed specificity of the 2‐oxo acid dehydrogenase components of both complexes for these lipoyl domains is discussed in this respect.Keywords
This publication has 49 references indexed in Scilit:
- Three-dimensional structure of a lipoyl domain fromthe dihydrolipoyl acetyltransferase component of the pyruvate dehydrogenase multienzyme complex of Escherichia coliJournal of Molecular Biology, 1995
- Structural Dependence of Post-translational Modification and Reductive Acetylation of the Lipoyl Domain of the Pyruvate Dehydrogenase Multienzyme ComplexJournal of Molecular Biology, 1994
- Three-dimensional Structure of Lipoamide Dehydrogenase from Pseudomonas fluorescens at 2·8 Å Resolution: Analysis of Redox and Thermostability PropertiesJournal of Molecular Biology, 1993
- Refined Crystal Structure of the Catalytic Domain of Dihydrolipoyl Transacetylase (E2p) from Azotobacter vinelandii at 2·6 Å ResolutionJournal of Molecular Biology, 1993
- The High-resolution Structure of the Peripheral Subunit-binding Domain of Dihydrolipoamide Acetyltransferase from the Pyruvate Dehydrogenase Multienzyme Complex of Bacillus stearothermophilusJournal of Molecular Biology, 1993
- Relationship between nuclear magnetic resonance chemical shift and protein secondary structureJournal of Molecular Biology, 1991
- Refined crystal structure of lipoamide dehydrogenase from Azotobacter vinelandii at 2.2 Å resolutionJournal of Molecular Biology, 1991
- X-ray structure of lipoamide dehydrogenase from Azotobacter vinelandii determined by a combination of molecular and isomorphous replacement techniquesJournal of Molecular Biology, 1989
- Improved spectral resolution in COSY 1H NMR spectra of proteins via double quantum filteringBiochemical and Biophysical Research Communications, 1983
- A two-dimensional nuclear Overhauser enhancement (2D NOE) experiment for the elucidation of complete proton-proton cross-relaxation networks in biological macromoleculesBiochemical and Biophysical Research Communications, 1980