Research article

Post-translational modifications of the apelin receptor regulate its functional expression

  • Received: 14 July 2023 Revised: 24 October 2023 Accepted: 26 October 2023 Published: 31 October 2023
  • Post-translational modifications (PTMs) are protein modifications that occur after protein biosynthesis, playing a crucial role in regulating protein function. They are involved in the functional expression of G-protein-coupled receptors (GPCRs), as well as intracellular and secretory protein signaling. Here, we aimed to investigate the PTMs of the apelin receptor (APLNR), a GPCR and their potential influence on the receptor's function. In an in vitro experiment using HEK cells, we only observed glycosylation as a PTM of the APLNR and ineffective receptor signaling by the agonist, (Pyr1)-apelin-13. In contrast, when analyzing mouse spinal cord, we detected glycosylation and other PTMs, excluding isopeptidation. This suggests that additional PTMs are involved in the functional expression of the APLNR in vitro. In summary, these findings suggest that the APLNR in vivo requires multiple PTMs for functional expression. To comprehensively understand the pharmacological effects of the APLNR, it is essential to establish an in vitro system that adequately replicates the receptor's PTM profile. Nonetheless, it is crucial to overcome the challenge of heat-sensitive proteolysis in APLNR studies. By elucidating the regulation of PTMs, further research has the potential to advance the analysis and pharmacological studies of both the apelin/APLNR system and GPCR signal modulation.

    Citation: Toshihiko Kinjo, Shun Ebisawa, Tatsuya Nokubo, Mifu Hashimoto, Takonori Yamada, Michiko Oshio, Ruka Nakamura, Kyosuke Uno, Nobuyuki Kuramoto. Post-translational modifications of the apelin receptor regulate its functional expression[J]. AIMS Neuroscience, 2023, 10(4): 282-299. doi: 10.3934/Neuroscience.2023022

    Related Papers:

  • Post-translational modifications (PTMs) are protein modifications that occur after protein biosynthesis, playing a crucial role in regulating protein function. They are involved in the functional expression of G-protein-coupled receptors (GPCRs), as well as intracellular and secretory protein signaling. Here, we aimed to investigate the PTMs of the apelin receptor (APLNR), a GPCR and their potential influence on the receptor's function. In an in vitro experiment using HEK cells, we only observed glycosylation as a PTM of the APLNR and ineffective receptor signaling by the agonist, (Pyr1)-apelin-13. In contrast, when analyzing mouse spinal cord, we detected glycosylation and other PTMs, excluding isopeptidation. This suggests that additional PTMs are involved in the functional expression of the APLNR in vitro. In summary, these findings suggest that the APLNR in vivo requires multiple PTMs for functional expression. To comprehensively understand the pharmacological effects of the APLNR, it is essential to establish an in vitro system that adequately replicates the receptor's PTM profile. Nonetheless, it is crucial to overcome the challenge of heat-sensitive proteolysis in APLNR studies. By elucidating the regulation of PTMs, further research has the potential to advance the analysis and pharmacological studies of both the apelin/APLNR system and GPCR signal modulation.


    Abbreviations

    AKT

    protein kinase B

    APLNR

    apelin receptor

    ATG

    autophagy-related protein

    cAMP

    cyclic adenosine monophosphate

    ERK

    extracellular signal regulated kinase

    FAT 10

    F locus adjacent transcript 10

    FSK

    forskolin

    GFP

    green Fluorescent Protein

    GPCR

    G-protein coupled receptor

    GPI

    glycosylphosphatidylinositol

    Gpr 44

    G protein-coupled receptor 44

    HEK

    human embryonic kidney

    ISG 15

    interferon-stimulated gene 15

    LPA 1

    lysophosphatidic acid receptor 1

    MAPK

    mitogen-activated protein kinase

    mTOR

    mammalian target of rapamycin

    NEDD

    neural-precursor-cell-expressed developmentally down-regulated protein

    NMDA

    N-methyl-D-aspartate

    PAR-1

    protease-activated receptor 1

    PDK1

    phosphoinositide-dependent protein kinase 1

    PI3K

    phosphoinositide 3-kinase

    PKA

    protein kinase A

    PTM

    post-translational modification

    SDS-PAGE

    sodium dodecyl sulfate polyacrylamide gel electrophoresis

    SUMO

    small ubiquitin-like modifier

    UFM 1

    ubiquitin-fold modifier

    URM 1

    ubiquitin-related modifier-1

    加载中

    Acknowledgments



    This work was supported by the Grant-in-Aid for Scientific Research from the Japan Society for the Promotion of Science (JSPS), Ministry of Education, Culture, Sports, Science and Technology of Japan (Grant Number JP21K06681).

    Conflict of interest



    The authors declare no conflict of interest.

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