title: Conformations of JNK3α splice variants analyzed by hydrogen/deuterium exchange mass spectrometry.
abstract: c-Jun N-terminal kinases (JNKs) are members of the mitogen-activated protein kinase (MAPK) family that regulate apoptosis, inflammation, cytokine production, and metabolism. MAPKs undergo various splicing within their kinase domains. Unlike other MAPKs, JNKs have alternative splicing at the C-terminus, resulting in long and short variants. Functional or conformational effects due to the elongated C-terminal tail in the long splice variants have not been investigated nor has the conformation of the C-terminal tail been analyzed. Here, we analyzed the conformation of the elongated C-terminal tail and investigated conformational differences between long and short splice variants of JNKs using JNK3α2 and JNK3α1 as models. We adopted hydrogen/deuterium exchange mass spectrometry (HDX-MS) to analyze the conformation. HDX-MS revealed that the C-terminal tail is mostly intrinsically disordered, and that the conformation of the kinase domain of JNK3α2 is more dynamic than that of JNK3α1. The different conformation dynamics between long and short splice variants of JNK3α might affect the cellular functions of JNK3.
authors.affiliation: School of Pharmacy, Sungkyunkwan University, Suwon, Republic of Korea.
authors.forename: Ji Young
authors.initials: JY
authors.lastname: Park
authors.affiliation: School of Pharmacy, Sungkyunkwan University, Suwon, Republic of Korea.
authors.forename: Youngjoo
authors.initials: Y
authors.lastname: Yun
authors.affiliation: School of Pharmacy, Sungkyunkwan University, Suwon, Republic of Korea. Electronic address: kychung2@skku.edu.
authors.forename: Ka Young
authors.initials: KY
authors.lastname: Chung
mesh_terms: D017398:Alternative Splicing / Q000235:genetics; D000595:Amino Acid Sequence; D003903:Deuterium / Q000737:chemistry; D041961:Deuterium Exchange Measurement / Q000379:methods*; D006801:Humans; D006859:Hydrogen / Q000737:chemistry; D013058:Mass Spectrometry / Q000379:methods*; D048057:Mitogen-Activated Protein Kinase 10 / Q000737:chemistry* / Q000235:genetics*; D011487:Protein Conformation; D017433:Protein Structure, Secondary; D012326:RNA Splicing / Q000235:genetics; D017386:Sequence Homology, Amino Acid
chemical_list: D006859:Hydrogen; D003903:Deuterium; D048057:Mitogen-Activated Protein Kinase 10
keywords: Conformation; HDX-MS; JNK; Splice variant