Departamento de Biología Celular y Desarrollo, Ap. Postal 70-600, Ciudad de México, 04510, Mexico.
Unidad de Imagenología, Instituto de Fisiología Celular, Universidad Nacional Autónoma de México, Ciudad Universitaria, Ap. Postal 70-600, Ciudad de México, 04510, Mexico.
Mol Cell Endocrinol. 2023 Jun 15;570:111930. doi: 10.1016/j.mce.2023.111930. Epub 2023 Apr 11.
LPA internalization to endosomes was studied employing Förster Resonance Energy Transfer (FRET) in cells coexpressing the mCherry-lysophosphatidic acid LPA receptors and distinct eGFP-tagged Rab proteins. Lysophosphatidic acid (LPA)-induced internalization was rapid and decreased afterward: phorbol myristate acetate (PMA) action was slower and sustained. LPA stimulated LPA-Rab5 interaction rapidly but transiently, whereas PMA action was rapid but sustained. Expression of a Rab5 dominant-negative mutant blocked LPA-Rab5 interaction and receptor internalization. LPA-induced LPA-Rab9 interaction was only observed at 60 min, and LPA-Rab7 interaction after 5 min with LPA and after 60 min with PMA. LPA triggered immediate but transient rapid recycling (i.e., LPA-Rab4 interaction), whereas PMA action was slower but sustained. Agonist-induced slow recycling (LPA-Rab11 interaction) increased at 15 min and remained at this level, whereas PMA action showed early and late peaks. Our results indicate that LPA receptor internalization varies with the stimuli.
采用共表达 mCherry-溶血磷脂酸（LPA）受体和不同的 eGFP 标记 Rab 蛋白的细胞中的Förster 共振能量转移（FRET）研究了 LPA 向内体的内化。溶血磷脂酸（LPA）诱导的内化迅速发生，随后减少：佛波醇 12,13-二丁酸酯（PMA）的作用较慢且持续。LPA 快速但短暂地刺激 LPA-Rab5 相互作用，而 PMA 的作用则快速但持续。表达 Rab5 显性负突变体阻断了 LPA-Rab5 相互作用和受体内化。仅在 60 分钟时观察到 LPA 诱导的 LPA-Rab9 相互作用，而在 5 分钟时用 LPA 观察到 LPA-Rab7 相互作用，在 60 分钟时用 PMA 观察到 LPA-Rab7 相互作用。LPA 触发立即但短暂的快速再循环（即，LPA-Rab4 相互作用），而 PMA 的作用较慢但持续。激动剂诱导的缓慢再循环（LPA-Rab11 相互作用）在 15 分钟时增加并保持在此水平，而 PMA 的作用显示早期和晚期峰值。我们的结果表明，LPA 受体内化随刺激而变化。
