title: Preparation of functional human lysophosphatidic acid receptor 2 using a P9∗expression system and an amphipathic polymer and investigation of its in vitro binding preference to Gαproteins.
abstract: Human lysophosphatidic acid receptor 2 (LPA2), a member of the G-protein coupled receptor family, mediates lysophosphatidic acid (LPA)-dependent signaling by recruiting various G proteins. Particularly, it is directly implicated in the progression of colorectal and ovarian cancer through G protein signaling cascades. To investigate the biochemical binding properties of LPA2against various alpha subunits of G protein (Gα), a functional recombinant LPA2was overexpressed in E. coli membrane with a P9∗expression system, and the purified protein was stabilized with an amphipathic polymer that had been synthesized by coupling octylamine, glucosamine, and diethyl aminoproylamine at the carboxylic groups of poly-γ-glutamic acid. The purified LPA2stabilized with the amphipathic polymer showed selective binding activity to the various Gαproteins as well as agonist-dependent dissociation from Gαi3. Understanding the binding properties of LPA2against various Gαproteins advances the understanding of downstream signaling cascades of LPA2. The functional LPA2prepared using a P9∗expression system and an amphipathic polymer could also facilitate the development of LPA2-targeting drugs.
authors.affiliation: Department of Chemistry, Kookmin University, 861-1 Jeongneung-dong, Seongbuk-gu, Seoul 136-702, South Korea.
authors.forename: Seong-Gu
authors.initials: SG
authors.lastname: Han
authors.affiliation: Department of Chemistry, Kookmin University, 861-1 Jeongneung-dong, Seongbuk-gu, Seoul 136-702, South Korea.
authors.forename: Seung-Il
authors.initials: SI
authors.lastname: Baek
authors.affiliation: Department of Chemistry, Kookmin University, 861-1 Jeongneung-dong, Seongbuk-gu, Seoul 136-702, South Korea.
authors.forename: Tae Jin
authors.initials: TJ
authors.lastname: Son
authors.affiliation: Department of Chemistry, Kookmin University, 861-1 Jeongneung-dong, Seongbuk-gu, Seoul 136-702, South Korea.
authors.forename: Hyeongjin
authors.initials: H
authors.lastname: Lee
authors.affiliation: Department of Chemistry, Kookmin University, 861-1 Jeongneung-dong, Seongbuk-gu, Seoul 136-702, South Korea.
authors.forename: Nam Hyuk
authors.initials: NH
authors.lastname: Kim
authors.affiliation: Department of Chemistry, Kookmin University, 861-1 Jeongneung-dong, Seongbuk-gu, Seoul 136-702, South Korea. Electronic address: ygyu@kookmin.ac.kr.
authors.forename: Yeon Gyu
authors.initials: YG
authors.lastname: Yu
mesh_terms: D001665:Binding Sites; D003001:Cloning, Molecular / Q000379:methods; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism*; D044385:GTP-Binding Protein alpha Subunits / Q000737:chemistry*; D006801:Humans; D004860:Infectious Anemia Virus, Equine / Q000235:genetics*; D011485:Protein Binding; D049368:Receptors, Lysophosphatidic Acid / Q000737:chemistry* / Q000502:physiology*
chemical_list: D044385:GTP-Binding Protein alpha Subunits; D049368:Receptors, Lysophosphatidic Acid
keywords: G-protein; GPCR; Lysophosphatidic acid receptor 2