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Membrane Restructuring Events during the Enzymatic Generation of Ceramides with Very Long-Chain Polyunsaturated Fatty Acids
Langmuir ( IF 3.7 ) Pub Date : 2018-03-15 00:00:00 , DOI: 10.1021/acs.langmuir.7b04374 Daniel A. Peñalva 1, 2 , Silvia S. Antollini 1, 2 , Ernesto E. Ambroggio 3, 4 , Marta I. Aveldaño 1, 2 , María L. Fanani 3, 4
Langmuir ( IF 3.7 ) Pub Date : 2018-03-15 00:00:00 , DOI: 10.1021/acs.langmuir.7b04374 Daniel A. Peñalva 1, 2 , Silvia S. Antollini 1, 2 , Ernesto E. Ambroggio 3, 4 , Marta I. Aveldaño 1, 2 , María L. Fanani 3, 4
Affiliation
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In rat sperm heads, sphingomyelin (SM) species that contain very long-chain polyunsaturated fatty acid (V-SM) become ceramides (V-Cer) after inducing in vitro the acrosomal reaction. The reason for such a specific location of this conversion, catalyzed by a sphingomyelinase (SMase), has received little investigation so far. Here, the effects of SMase were compared in unilamellar vesicles (large unilamellar vesicles (LUVs), giant unilamellar vesicles (GUVs)) containing phosphatidylcholine, and either V-SM or a palmitate-rich SM (P-SM). In uniformly sized LUVs at 37 °C, more V-Cer was generated and more rapidly than P-Cer. Nephelometry and dynamic light scattering showed that LUVs tended to form large lipid particles more intensely, and Förster resonance energy transfer (FRET) increases suggested that lateral lipid mixing was more marked when V-Cer rather than P-Cer was produced. As reported by 6-dodecanoyl-2-dimethyl-aminopnaphthalene (Laurdan) and 1,6-diphenyl-1,3,5,-hexatriene (DPH), the production of V-Cer resulted in higher and faster restriction in lipid mobility than that of P-Cer, implying a stronger increase in membrane dehydration and microviscosity. Moreover, DPH anisotropy suggested a higher solubility of V-Cer than that of P-Cer in the liquid-disordered phase. At room temperature, liquid-condensed lateral domains appeared in P-SM- but not in V-SM-containing GUVs. The former maintained their size while losing their contents gradually during SMase action, whereas the latter became permeable earlier and reduced their size in few minutes until suddenly collapsing. The fast and potent generation of V-Cer may contribute to the membrane restructuring events that occur on the acrosome-reacted sperm head.
中文翻译：
用很长链的多不饱和脂肪酸酶促生成神经酰胺过程中的膜重构事件
在大鼠精子头中，含有极长链多不饱和脂肪酸（V-SM）的鞘磷脂（SM）物种在体外诱导顶体反应后变为神经酰胺（V-Cer）。迄今为止，由鞘磷脂酶（SMase）催化的这种转化的这种特定位置的原因尚未得到研究。在这里，比较了SMase在含磷脂酰胆碱和V-SM或富含棕榈酸酯的SM（P-SM）的单层囊泡（大单层囊泡（LUV），巨型单层囊泡（GUV））中的作用。在37°C的大小均等的LUV中，生成的V-Cer比P-Cer更快。比浊法和动态光散射表明，LUV倾向于更强烈地形成大脂质颗粒，Förster共振能量转移（FRET）的增加表明，当生产V-Cer而不是P-Cer时，侧向脂质混合更为明显。如6-十二烷酰基-2-二甲基-氨基萘（Laurdan）和1,6-二苯基-1,3,5，-己三烯（DPH）所报道，V-Cer的产生比脂质迁移率的限制更高和更快。与P-Cer相比，意味着膜脱水和微粘度的增加更大。而且，DPH各向异性表明，V-Cer在液相无序相中的溶解度高于P-Cer。在室温下，液体冷凝的侧向区域出现在含P-SM的GUV中，而没有出现在含V-SM的GUV中。前者在SMase作用期间保持其大小，同时逐渐失去其内含物，而后者则较早变得可渗透，并在几分钟内减小了大小，直到突然崩溃。
更新日期：2018-03-15
中文翻译：
用很长链的多不饱和脂肪酸酶促生成神经酰胺过程中的膜重构事件
在大鼠精子头中，含有极长链多不饱和脂肪酸（V-SM）的鞘磷脂（SM）物种在体外诱导顶体反应后变为神经酰胺（V-Cer）。迄今为止，由鞘磷脂酶（SMase）催化的这种转化的这种特定位置的原因尚未得到研究。在这里，比较了SMase在含磷脂酰胆碱和V-SM或富含棕榈酸酯的SM（P-SM）的单层囊泡（大单层囊泡（LUV），巨型单层囊泡（GUV））中的作用。在37°C的大小均等的LUV中，生成的V-Cer比P-Cer更快。比浊法和动态光散射表明，LUV倾向于更强烈地形成大脂质颗粒，Förster共振能量转移（FRET）的增加表明，当生产V-Cer而不是P-Cer时，侧向脂质混合更为明显。如6-十二烷酰基-2-二甲基-氨基萘（Laurdan）和1,6-二苯基-1,3,5，-己三烯（DPH）所报道，V-Cer的产生比脂质迁移率的限制更高和更快。与P-Cer相比，意味着膜脱水和微粘度的增加更大。而且，DPH各向异性表明，V-Cer在液相无序相中的溶解度高于P-Cer。在室温下，液体冷凝的侧向区域出现在含P-SM的GUV中，而没有出现在含V-SM的GUV中。前者在SMase作用期间保持其大小，同时逐渐失去其内含物，而后者则较早变得可渗透，并在几分钟内减小了大小，直到突然崩溃。
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