[PMID]: | 28283404 |
[Au] Autor: | Rangl M; Rima L; Klement J; Miyagi A; Keller S; Scheuring S |
[Ad] Endereço: | Department of Anesthesiology, Physiology and Biophysics, Weill Cornell Medical College, 1300 York Avenue, New York, NY 10065, USA; INSERM U1006, Aix-Marseille Université, Parc Scientifique et Technologique de Luminy, 163 Avenue de Luminy, 13009 Marseille, France. |
[Ti] Título: | Real-time Visualization of Phospholipid Degradation by Outer Membrane Phospholipase A using High-Speed Atomic Force Microscopy. |
[So] Source: | J Mol Biol;429(7):977-986, 2017 Apr 07. |
[Is] ISSN: | 1089-8638 |
[Cp] País de publicação: | England |
[La] Idioma: | eng |
[Ab] Resumo: | Phospholipases are abundant in various types of cells and compartments, where they play key roles in physiological processes as diverse as digestion, cell proliferation, and neural activation. In Gram-negative bacteria, outer membrane phospholipase A (OmpLA) is involved in outer-membrane lipid homeostasis and bacterial virulence. Although the enzymatic activity of OmpLA can be probed with an assay relying on an artificial monoacyl thioester substrate, only little is known about its activity on diacyl phospholipids. Here, we used high-speed atomic force microscopy (HS-AFM) to directly image enzymatic phospholipid degradation by OmpLA in real time. In the absence of Ca , reconstituted OmpLA diffused within a phospholipid bilayer without revealing any signs of phospholipase activity. Upon the addition of Ca , OmpLA was activated and degraded the membrane with a turnover of ~2 phospholipid molecules per second and per OmpLA dimer until most of the membrane phospholipids were hydrolyzed and the protein became tightly packed. |
[Mh] Termos MeSH primário: |
Proteínas da Membrana Bacteriana Externa/metabolismo Microscopia de Força Atômica/métodos Fosfolipases A1/metabolismo Fosfolipídeos/metabolismo
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[Mh] Termos MeSH secundário: |
Cálcio/metabolismo Cinética Modelos Biológicos
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[Pt] Tipo de publicação: | JOURNAL ARTICLE |
[Nm] Nome de substância:
| 0 (Bacterial Outer Membrane Proteins); 0 (Phospholipids); EC 3.1.1.32 (Phospholipases A1); EC 3.1.1.32 (outer membrane phospholipase A); SY7Q814VUP (Calcium) |
[Em] Mês de entrada: | 1707 |
[Cu] Atualização por classe: | 170703 |
[Lr] Data última revisão:
| 170703 |
[Sb] Subgrupo de revista: | IM |
[Da] Data de entrada para processamento: | 170312 |
[St] Status: | MEDLINE |
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