Group X secreted phospholipase A2 proenzyme is matured by a furin-like proprotein convertase and releases arachidonic acid inside of human HEK293 cells
| Autor: | Christine Payré, Khaoula Chargui, Rob C. Oslund, Sabine Scarzello, Michael H. Gelb, Hiromi, Gérard Lambeau, Anne Sophie Dabert-Gay, Dominique Douguet, Ikram Jemel |
|---|---|
| Přispěvatelé: | Institut de pharmacologie moléculaire et cellulaire (IPMC), Centre National de la Recherche Scientifique (CNRS)-Université Nice Sophia Antipolis (... - 2019) (UNS), COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-COMUE Université Côte d'Azur (2015-2019) (COMUE UCA)-Université Côte d'Azur (UCA), Department of Chemistry, University of Washington [Seattle], Department of Biochemistry [Washington ] |
| Jazyk: | angličtina |
| Rok vydání: | 2011 |
| Předmět: |
MESH: Group X Phospholipases A2
MESH: Mutation medicine.medical_treatment Amino Acid Motifs Biochemistry 03 medical and health sciences Mice MESH: Amino Acid Motifs MESH: Enzyme Precursors 0302 clinical medicine Phospholipase A2 medicine Animals Group X Phospholipases A2 Humans Secretion Protease Inhibitors MESH: Animals Protein precursor Molecular Biology Furin MESH: Mice 030304 developmental biology chemistry.chemical_classification 0303 health sciences Enzyme Precursors Protease Arachidonic Acid MESH: Protease Inhibitors MESH: Humans biology Cell Biology Proprotein convertase Lipids Amino acid MESH: Arachidonic Acid HEK293 Cells chemistry 030220 oncology & carcinogenesis MESH: HEK293 Cells Mutation biology.protein MESH: Proprotein Convertases [SDV.IMM]Life Sciences [q-bio]/Immunology Proprotein Convertases |
| Zdroj: | Journal of Biological Chemistry Journal of Biological Chemistry, American Society for Biochemistry and Molecular Biology, 2011, 286 (42), pp.36509-21. ⟨10.1074/jbc.M111.268540⟩ |
| ISSN: | 0021-9258 1083-351X |
| Popis: | International audience; Among mammalian secreted phospholipases A(2) (sPLA(2)s), group X sPLA(2) has the most potent hydrolyzing activity toward phosphatidylcholine and is involved in arachidonic acid (AA) release. Group X sPLA(2) is produced as a proenzyme and contains a short propeptide of 11 amino acids ending with a dibasic motif, suggesting cleavage by proprotein convertases. Although the removal of this propeptide is clearly required for enzymatic activity, the cellular location and the protease(s) involved in proenzyme conversion are unknown. Here we have analyzed the maturation of group X sPLA(2) in HEK293 cells, which have been extensively used to analyze sPLA(2)-induced AA release. Using recombinant mouse (PromGX) and human (ProhGX) proenzymes; HEK293 cells transfected with cDNAs coding for full-length ProhGX, PromGX, and propeptide mutants; and various permeable and non-permeable sPLA(2) inhibitors and protease inhibitors, we demonstrate that group X sPLA(2) is mainly converted intracellularly and releases AA before externalization from the cell. Most strikingly, the exogenous proenzyme does not elicit AA release, whereas the transfected proenzyme does elicit AA release in a way insensitive to non-permeable sPLA(2) inhibitors. In transfected cells, a permeable proprotein convertase inhibitor, but not a non-permeable one, prevents group X sPLA(2) maturation and partially blocks AA release. Mutations at the dibasic motif of the propeptide indicate that the last basic residue is required and sufficient for efficient maturation and AA release. All together, these results argue for the intracellular maturation of group X proenzyme in HEK293 cells by a furin-like proprotein convertase, leading to intracellular release of AA during secretion. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|