title: Purification and biophysical characterization of the AIMP2-DX2 protein.
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.
authors.affiliation: College of Pharmacy, Korea University, 2511 Sejong-ro, Sejong 30019, Republic of Korea.
authors.forename: Roshan
authors.initials: R
authors.lastname: Jha
authors.affiliation: College of Pharmacy, Korea University, 2511 Sejong-ro, Sejong 30019, Republic of Korea.
authors.forename: Hye Young
authors.initials: HY
authors.lastname: Cho
authors.affiliation: College of Pharmacy, Korea University, 2511 Sejong-ro, Sejong 30019, Republic of Korea.
authors.forename: Ameeq Ul
authors.initials: AU
authors.lastname: Mushtaq
authors.affiliation: College of Pharmacy, Korea University, 2511 Sejong-ro, Sejong 30019, Republic of Korea.
authors.forename: Kiho
authors.initials: K
authors.lastname: Lee
authors.affiliation: Medicinal Bioconvergence Research Center, Seoul National University, Seoul 08826, Republic of Korea; Research Institute of Pharmaceutical Sciences, College of Pharmacy, Seoul National University, Seoul 08826, Republic of Korea.
authors.forename: Dae Gyu
authors.initials: DG
authors.lastname: Kim
authors.affiliation: Medicinal Bioconvergence Research Center, Seoul National University, Seoul 08826, Republic of Korea; Research Institute of Pharmaceutical Sciences, College of Pharmacy, Seoul National University, Seoul 08826, Republic of Korea.
authors.forename: Sunghoon
authors.initials: S
authors.lastname: Kim
authors.affiliation: College of Pharmacy, Korea University, 2511 Sejong-ro, Sejong 30019, Republic of Korea. Electronic address: yhjeon@korea.ac.kr.
authors.forename: Young Ho
authors.initials: YH
authors.lastname: Jeon
mesh_terms: D006801:Humans; D019906:Nuclear Magnetic Resonance, Biomolecular; D009687:Nuclear Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D055503:Protein Multimerization; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D013329:Structure-Activity Relationship
chemical_list: C575685:AIMP2 protein, human; D009687:Nuclear Proteins; D011993:Recombinant Fusion Proteins
keywords: AIMP2; DX2; Dimer; NMR; Oncogenic