Purification and biophysical characterization of the AIMP2-DX2 protein

Roshan Jha, Hye Young Cho, Ameeq Ul Mushtaq, Kiho Lee, Dae Gyu Kim, Sunghoon Kim, Young Ho Jeon

Research output: Contribution to journalArticlepeer-review

3 Citations (Scopus)

Abstract

Besides their primary role in protein synthesis, aminoacyl-tRNA synthetases (AARSs) are involved in several non-canonical processes such as apoptosis, inflammation and angiogenesis through their interactions with various cellular proteins. Nine of these AARSs interact with three aminoacyl-tRNA synthetase interacting multifunctional proteins (AIMPs), forming a multi-synthetase complex (MSC) in eukaryotes. Among the three AIMPs, AIMP2 is involved in controlling cell proliferation and apoptosis. However, a splicing variant of AIMP2 lacking exon 2, referred to as AIMP2-DX2, is oncogenic and compromises the pro-apoptotic activity of AIMP2 by competing with it for p53 and TRAF2. AIMP2-DX2 is also an inhibitor of p14arf activity. Thus, there is a pressing need for structural insight into the oncogenic role of AIMP2-DX2. In this study, we expressed and purified human AIMP2-DX2 using a SUMO tag to more than 95% purity and a yield of 10 mg/L. We have used size exclusion chromatography, glutaraldehyde cross-linking, dynamic light scattering and nuclear magnetic resonance spectroscopy to characterize its biophysical properties. These data indicate monomer-dimer equilibrium of AIMP2-DX2 in solution. These results form the basis for the structure-function study of oncogenic AIMP2-DX2.

Original languageEnglish
Pages (from-to)131-137
Number of pages7
JournalProtein Expression and Purification
Volume132
DOIs
Publication statusPublished - 2017 Apr 1

Bibliographical note

Funding Information:
This work was supported by the Global Frontier Project (grants NRF-M1AXA002-20100029785 (S.K.) and NRF-2013M3A6A4045160 and 2014R1A4A1007304 (Y.H.J.) of the National Research Foundation funded by the Ministry of Science, ICT & Future Planning (MSIP) of Korea.

All Science Journal Classification (ASJC) codes

  • Biotechnology

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