Quantitative multiple reaction monitoring analysis of synaptic proteins from human brain

Chang, Rachel Yoon Kyung, Etheridge, Naomi, Dodd, Peter and Nouwens, Amanda (2014) Quantitative multiple reaction monitoring analysis of synaptic proteins from human brain. Journal of Neuroscience Methods, 227 189-210. doi:10.1016/j.jneumeth.2014.02.016


Author Chang, Rachel Yoon Kyung
Etheridge, Naomi
Dodd, Peter
Nouwens, Amanda
Title Quantitative multiple reaction monitoring analysis of synaptic proteins from human brain
Journal name Journal of Neuroscience Methods   Check publisher's open access policy
ISSN 0165-0270
1872-678X
Publication date 2014-04-30
Year available 2014
Sub-type Article (original research)
DOI 10.1016/j.jneumeth.2014.02.016
Volume 227
Start page 189
End page 210
Total pages 22
Place of publication Amsterdam, Netherlands
Publisher Elsevier
Language eng
Subject 2800 Neuroscience
Formatted abstract
Background: The recent introduction of multiple reaction monitoring to proteomics research has allowed many researchers to apply this technique to study human diseases.

New methods: Here we combine subcellular fractionation of human autopsy brain with label-free multiple reaction monitoring to quantitatively analyse proteins in synapses. The protein enolase, from Streptococcus pyogenes serotype M6, which is sufficiently different from human proteins, was spiked into the sample mixture prior to trypsin digestion and used as an internal standard across samples.

Results: Three synaptic proteins and an internal standard analysed with four injections over four consecutive days gave consistent differences with a coefficient of variation of <4%. Consistent retention time was recorded across the replicates. Comparison with existing methods: Previously, multiple reaction monitoring analysis has been utilized to study human autopsy and animal tissues. Utilizing the synaptosomal fraction prior to analysis reduced sample complexity and allowed the enriched synaptic proteins to be quantitatively assessed in a highly reproducible manner, without the need for expensive fluorescent labels and synthetic peptides.

Conclusion: Protein expression can be measured with accuracy using label-free multiple reaction monitoring mass spectrometry in relatively complex human brain samples. Synaptic functions are critical for neuronal communication and function, and synapse dysfunction underlies many neurodegenerative diseases, including Alzheimer's disease. This method can be applied to study a range of brain disorders.
Keyword Alzheimer's disease
Human autopsy brain
Neurodegeneration
Sample enrichment
Synapse
Q-Index Code C1
Q-Index Status Confirmed Code
Institutional Status UQ

Document type: Journal Article
Sub-type: Article (original research)
Collections: Official 2015 Collection
School of Chemistry and Molecular Biosciences
 
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