Native Piezo2 Interactomics Identifies Pericentrin as a Novel Regulator of Piezo2 in Somatosensory Neurons
† Max-Planck
Institute of Experimental Medicine, Somatosensory
Signaling and Systems Biology Group, D-37075 Goettingen, Germany
‡ Max
Planck Institute of Biophysical Chemistry, Bioanalytical Mass Spectrometry Group, D-37077 Goettingen, Germany
§ Bioanaytics
Group, Institute for Clinical Chemistry, University Medical Center Göttingen, D-37075 Göttingen, Germany
∥ Max-Planck
Institute of Experimental Medicine, Light
Microscopy Facility, D-37075 Goettingen, Germany
J. Proteome Res., 2016, 15 (8), pp 2676–2687
DOI: 10.1021/acs.jproteome.6b00235
Publication Date (Web): June 26, 2016
Copyright © 2016 American Chemical Society
*D.G.-V.: Tel: 49-551-3899-572. Fax: 49-551-3899-573. E-mail: gomezvarela@em.mpg.de., *M.S.: Tel: 49-551-3899-572. Fax: 49-551-3899-573. E-mail: mschmidt@em.mpg.de.
Abstract
The ability of somatosensory neurons to perceive mechanical stimuli relies on specialized mechanotransducing proteins and their molecular environment. Only recently has the identity of a major transducer of mechanical forces in vertebrates been revealed by the discovery of Piezo2. Further work has established its pivotal role for innocuous touch in mice. Therefore, Piezo2 offers a unique platform for the molecular investigation of somatosensory mechanosensation. We performed a mass spectrometry-based interactomics screen on native Piezo2 in somatosensory neurons of mouse dorsal root ganglia (DRG). Stringent and quantitative data analysis yielded the identity of 36 novel binding partners of Piezo2. The biological significance of this data set is reflected by functional experiments demonstrating a role for Pericentrin in modulating Piezo2 activity and membrane expression in somatosensory neurons. Collectively, our findings provide a framework for understanding Piezo2 physiology and serve as a rich resource for the molecular dissection of mouse somatosensation.
The Supporting Information is available free of charge on the ACS Publications website at DOI: 10.1021/acs.jproteome.6b00235.
Figure S-1: Specificity test of commercial Piezo2 antibodies. Figure S-2: Piezo2-immunoaffinity-purification with Piezo2-Ab1 compared to controls. Figure S-3: Co-immunoprecipitation of Piezo2 and Pcnt in HEK293T cells. Figure S-4: Piezo2-currents upon coexpression with Pcnt in HEK293T cells. Figure S-5: Specificity of Pcnt antibodies. Figure S-6: Pcnt expression in lanceolate endings. (PDF)
Table S-1: Results of quantitative mass spectrometry from all replicates of Piezo2-affinity-purfications compared to controls. (XLS)
