Steady-State Kinetics of α-Synuclein Ferrireductase Activity Identifies the Catalytically Competent Species

Jennifer S McDowall, Ioanna Ntai, Jonathon Hake, Paul R Whitley, Jody M Mason, Christopher R Pudney, David R Brown

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Abstract

α-Synuclein (α-syn) is a cytosolic protein known for its association with neurodegenerative diseases, including Parkinson's disease and other synucleinopathies. The potential cellular function of α-synuclein may be of consequence for understanding the pathogenesis of such diseases. Previous work has suggested that α-synuclein can catalyze the reduction of iron as a ferrireductase. We performed a detailed analysis of the steady-state kinetics of recombinant α-syn ferrireductase activity and for disease-associated variants. Our study illustrates that the ferrireductase activity we observed is clearly commensurate with bona fide enzyme activity and suggests a mechanistic rationale for the activity and the relationship to cellular regulation of the pool of Fe(III) and Fe(II). Using cell-based studies, we examined the functionally active conformation and found that the major catalytically active form is a putative membrane-associated tetramer. Using an artificial membrane environment with recombinant protein, we demonstrate that secondary structure folding of α-synuclein is insufficient to allow enzyme activity and the absolute specificity of the tertiary/quaternary structure is the primary requirement. Finally, we explored the steady-state kinetics of a range of disease α-synuclein variants and found that variants involved in neurodegenerative disease exhibited major changes in their enzymatic activity. We discuss these data in the context of a potential disease-associated mechanism for aberrant α-synuclein ferrireductase activity.

Original languageEnglish
Pages (from-to)2497–2505
Number of pages9
JournalBiochemistry
Volume56
Issue number19
Early online date4 May 2017
DOIs
Publication statusPublished - 16 May 2017

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alpha-Synuclein
Kinetics
Neurodegenerative diseases
Enzyme activity
Neurodegenerative Diseases
Artificial Membranes
Membranes
Enzymes
ferric citrate iron reductase
Recombinant Proteins
Parkinson Disease
Conformations
Iron
Association reactions

Keywords

  • Journal Article

Cite this

Steady-State Kinetics of α-Synuclein Ferrireductase Activity Identifies the Catalytically Competent Species. / McDowall, Jennifer S; Ntai, Ioanna; Hake, Jonathon; Whitley, Paul R; Mason, Jody M; Pudney, Christopher R; Brown, David R.

In: Biochemistry, Vol. 56, No. 19, 16.05.2017, p. 2497–2505.

Research output: Contribution to journalArticle

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