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Int J Physiol Pathophysiol Pharmacol 2012;4(2):74-83.

Original Article
Proteomic analysis of 14-3-3 zeta binding proteins in the mouse hippocampus

Maura Heverin, Gary P Brennan, Christian J Koehler, Achim Treumann, David C Henshall

Department of Physiology and Medical Physics; Molecular and Cellular Therapeutics, Royal College of Surgeons in Ireland, Dublin,

Received June 1, 2012; accepted June 20, 2012; Epub June 23, 2012; Published June 30, 2012

Abstract: 14-3-3 proteins are ubiquitous molecular chaperones with important roles in brain development and neuronal function.
Altered expression of 14-3-3 proteins has been reported in several neurologic and neurodegenerative disorders and identifying 14-3-3
binding proteins may provide important insights into the physiologic and pathophysiologic roles of these proteins. Particular interest
has emerged on 14-3-3 zeta (ζ) in the setting of neuronal injury because reducing 14-3-3ζ levels triggers an endoplasmic reticulum
stress-like response in neurons and increases vulnerability to excitotoxicity. Here we examined the subcellular distribution of 14-3-3
zeta (ζ) in the mouse hippocampus. We then used recombinant His-tagged 14-3-3ζ to pull-down interacting proteins from the mouse
hippocampus followed by identification by liquid chromatography-mass spectrometry. 14-3-3ζ protein was present in the cytoplasm,
microsomal compartment, nucleus and mitochondrial fractions of the mouse hippocampus. Recombinant 14-3-3ζ eluted 13 known 14-
3-3 binding partners, including three other 14-3-3 isoforms, and 16 other proteins which have not previously been reported to bind 14-3-
3ζ. The present study identifies potentially novel 14-3-3ζ binding proteins and contributes to defining the 14-3-3ζ interactome in the
mouse brain. (IJPPP1206001)

Keywords: 14-3-3 interactome, apoptosis, epilepsy, proteomics, mass spectrometry, hippocampus

Address all correspondence to:
Dr. David C. Henshall
Department of Physiology and Medical Physics
Royal College of Surgeons in Ireland
123 St. Stephen’s Green, Dublin 2, Ireland.
Tel: +353 1 402 8629; Fax: +353 1 402 2447
E-mail: dhenshall@rcsi.ie