@article{e2390ef3eff549849373f549c8e0ffe2,
title = "Mass spectrometry analysis of affinity-purified cytoplasmic translation initiation complexes from human and fly cells",
abstract = "eIF5-mimic protein (5MP) controls translation through binding to the ribosomal pre-initiation complex (PIC) and alters non-AUG translation rates for cancer oncogenes and repeat-expansions in neurodegenerative diseases. Here, we describe a semi-quantitative protocol for detecting 5MP-associated proteins in cultured human and fly cells. We detail one-step anti-FLAG affinity purification and whole-lane mass spectrometry analysis of samples resolved by SDS-PAGE. This protocol allows for quantitative evaluation of the effect of 5MP mutations on its molecular interactions, to elucidate translational control by 5MP. For complete details on the use and execution of this protocol, please refer to Singh et al. (2021).",
author = "Singh, {Chingakham Ranjit} and Naoki Tani and Akira Nakamura and Katsura Asano",
note = "Funding Information: This work was supported by a pilot grant from the K-INBRE Program P20 GM103418 , National Institutes of Health ; Innovative Research Awards from the Kansas State University (KSU) Terry Johnson Cancer Center ; National Science Foundation Research Grant (No. 1412250 ), National Institutes of Health grant GM125671 , the Joint Usage/ Research Center for Developmental Medicine , IMEG , Kumamoto University and JSPS International Collaboration Enhancement Grant 18K19963 (to K.A.). Funding Information: This work was supported by a pilot grant from the K-INBRE Program P20 GM103418, National Institutes of Health; Innovative Research Awards from the Kansas State University (KSU) Terry Johnson Cancer Center; National Science Foundation Research Grant (No. 1412250), National Institutes of Health grant GM125671, the Joint Usage/Research Center for Developmental Medicine, IMEG, Kumamoto University and JSPS International Collaboration Enhancement Grant 18K19963 (to K.A.). C.R.S. N.T. A.N. and K.A. wrote the original draft. K.A. and C.R.S. edited and completed the manuscript. Authors declare no conflict of interests. Publisher Copyright: {\textcopyright} 2022 The Author(s)",
year = "2022",
month = dec,
day = "16",
doi = "10.1016/j.xpro.2022.101739",
language = "English (US)",
journal = "STAR Protocols",
issn = "2666-1667",
publisher = "Cell Press",
}