posted on 2020-03-24, 13:06authored byYeon Choi, Kyowon Jeong, Sanghee Shin, Joon Won Lee, Young-suk Lee, Sangtae Kim, Sun Ah Kim, Jaehun Jung, Kwang Pyo Kim, V. Narry Kim, Jong-Seo Kim
Quantitative
proteomic platforms based on precursor intensity in
mass spectrometry (MS1-level) uniquely support in vivo metabolic labeling with superior quantification accuracy but suffer
from limited multiplexity (≤3-plex) and frequent missing quantities.
Here we present a new MS1-level quantification platform that allows
maximal multiplexing with high quantification accuracy and precision
for the given labeling scheme. The platform currently comprises 6-plex in vivo SILAC or in vitro diethylation
labeling with a dedicated algorithm and is also expandable to higher
multiplexity (e.g., nine-plex for SILAC). For complex samples with
broad dynamic ranges such as total cell lysates, our platform performs
highly accurately and free of missing quantities. Furthermore, we
successfully applied our method to measure protein synthesis rate
under heat shock response in human cells by 6-plex pulsed SILAC experiments,
demonstrating the unique biological merits of our in vivo platform to disclose translational regulations for cellular response
to stress.