Activity-dependent changes in synaptic protein complex composition are consistent in different detergents despite differential solubility

At the post-synaptic density (PSD), large protein complexes dynamically form and dissociate in response to synaptic activity, comprising the biophysical basis for learning and memory. The use of detergents to isolate the PSD and release its membrane-associated proteins complicates studies of these a...

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Bibliographic Details
Published in:	Scientific reports Vol. 9; no. 1; pp. 10890 - 15
Main Authors:	Lautz, Jonathan D., Gniffke, Edward P., Brown, Emily A., Immendorf, Karen B., Mendel, Ryan D., Smith, Stephen E. P.
Format:	Journal Article
Language:	English
Published:	London Nature Publishing Group UK 26-07-2019 Nature Publishing Group
Subjects:	631/378/2591 631/378/340 64 64/60 82 82/1 82/80 96 96/31 96/95 Animals Brain - metabolism Brain - pathology Chromatography, Gel Deoxycholic Acid - metabolism Detergents Detergents - metabolism Electrical Synapses - physiology Glutamic acid receptors (metabotropic) Humanities and Social Sciences Learning - physiology Memory - physiology Mice Molecular weight multidisciplinary Octoxynol - metabolism Post-Synaptic Density - chemistry Post-Synaptic Density - metabolism Protein composition Protein interaction Protein Interaction Maps Protein Multimerization Proteins ras GTPase-Activating Proteins - metabolism Science Science (multidisciplinary) Solubility Synaptic density
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Summary:	At the post-synaptic density (PSD), large protein complexes dynamically form and dissociate in response to synaptic activity, comprising the biophysical basis for learning and memory. The use of detergents to isolate the PSD and release its membrane-associated proteins complicates studies of these activity-dependent protein interaction networks, because detergents can simultaneously disrupt the very interactions under study. Despite widespread recognition that different detergents yield different experimental results, the effect of detergent on activity-dependent synaptic protein complexes has not been rigorously examined. Here, we characterize the effect of three detergents commonly used to study synaptic proteins on activity-dependent protein interactions. We first demonstrate that SynGAP-containing interactions are more abundant in 1% Deoxycholate (DOC), while Shank-, Homer- and mGluR5-containing interactions are more abundant in 1% NP-40 or Triton. All interactions were detected preferentially in high molecular weight complexes generated by size exclusion chromatography, although the detergent-specific abundance of proteins in high molecular weight fractions did not correlate with the abundance of detected interactions. Activity-dependent changes in protein complexes were consistent across detergent types, suggesting that detergents do not isolate distinct protein pools with unique behaviors. However, detection of activity-dependent changes is more or less feasible in different detergents due to baseline solubility. Collectively, our results demonstrate that detergents affect the solubility of individual proteins, but activity-dependent changes in protein interactions, when detectable, are consistent across detergent types.
ISSN:	2045-2322 2045-2322
DOI:	10.1038/s41598-019-46690-y