A multipronged approach unravels unprecedented protein–protein interactions in the human 2-oxoglutarate dehydrogenase multienzyme complex

Jieyu Zhou, Luying Yang, Oliver Ozohanics, Xu Zhang, Junjie Wang, A. Ambrus, Palaniappa Arjunan, Roman Brukh, Natalia S. Nemeria, William Furey, Frank Jordan

Research output: Article

2 Citations (Scopus)

Abstract

The human 2-oxoglutaric acid dehydrogenase complex (hOGDHc) plays a pivotal role in the tricarboxylic acid (TCA) cycle, and its diminished activity is associated with neurodegenerative diseases. The hOGDHc comprises three components, hE1o, hE2o, and hE3, and we recently reported functionally active E1o and E2o components, enabling studies on their assembly. No atomic-resolution structure for the hE2o component is currently available, so here we first studied the interactions in the binary subcomplexes (hE1o– hE2o, hE1o– hE3, and hE2o– hE3) to gain insight into the strength of their interactions and to identify the interaction loci in them. We carried out multiple physico-chemical studies, including fluorescence, hydrogen– deuterium exchange MS (HDX-MS), and chemical cross-linking MS (CL-MS). Our fluorescence studies suggested a strong interaction for the hE1o– hE2o subcomplex, but a much weaker interaction in the hE1o– hE3 subcomplex, and failed to identify any interaction in the hE2o– hE3 subcomplex. The HDX-MS studies gave evidence for interactions in the hE1o–hE2o and hE1o– hE3 subcomplexes comprising full-length components, identifying: (i) the N-terminal region of hE1o, in particular the two peptides 18YVEEM22 and 27ENPKSVHK-SWDIF39 as constituting the binding region responsible for the assembly of the hE1o with both the hE2o and hE3 components into hOGDHc, an hE1 region absent in available X-ray structures; and (ii) a novel hE2o region comprising residues from both a linker region and from the catalytic domain as being a critical region interacting with hE1o. The CL-MS identified the loci in the hE1o and hE2o components interacting with each other.

Original languageEnglish
Pages (from-to)19213-19227
Number of pages15
JournalJournal of Biological Chemistry
Volume293
Issue number50
DOIs
Publication statusPublished - jan. 1 2018

Fingerprint

Multienzyme Complexes
Ketoglutarate Dehydrogenase Complex
Oxidoreductases
Deuterium
Hydrogen
Fluorescence
Neurodegenerative diseases
Citric Acid Cycle
Neurodegenerative Diseases
Catalytic Domain
X-Rays
X rays
Peptides
alpha-ketoglutaric acid

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

A multipronged approach unravels unprecedented protein–protein interactions in the human 2-oxoglutarate dehydrogenase multienzyme complex. / Zhou, Jieyu; Yang, Luying; Ozohanics, Oliver; Zhang, Xu; Wang, Junjie; Ambrus, A.; Arjunan, Palaniappa; Brukh, Roman; Nemeria, Natalia S.; Furey, William; Jordan, Frank.

In: Journal of Biological Chemistry, Vol. 293, No. 50, 01.01.2018, p. 19213-19227.

Research output: Article

Zhou, J, Yang, L, Ozohanics, O, Zhang, X, Wang, J, Ambrus, A, Arjunan, P, Brukh, R, Nemeria, NS, Furey, W & Jordan, F 2018, 'A multipronged approach unravels unprecedented protein–protein interactions in the human 2-oxoglutarate dehydrogenase multienzyme complex', Journal of Biological Chemistry, vol. 293, no. 50, pp. 19213-19227. https://doi.org/10.1074/jbc.RA118.005432
Zhou, Jieyu ; Yang, Luying ; Ozohanics, Oliver ; Zhang, Xu ; Wang, Junjie ; Ambrus, A. ; Arjunan, Palaniappa ; Brukh, Roman ; Nemeria, Natalia S. ; Furey, William ; Jordan, Frank. / A multipronged approach unravels unprecedented protein–protein interactions in the human 2-oxoglutarate dehydrogenase multienzyme complex. In: Journal of Biological Chemistry. 2018 ; Vol. 293, No. 50. pp. 19213-19227.
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