Serotonylation of vascular proteins important to contraction.
Serotonylation of vascular proteins important to contraction.
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DOI：
10.1371/journal.pone.0005682
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发表时间：
2009-05-25
期刊：
影响因子：
3.7
通讯作者：
Thompson JM
中科院分区：
文献类型：
--
作者：
Watts SW;Priestley JR;Thompson JM
5-羟色胺（5-羟色胺，5-HT）因其来源（sero-）和改变平滑肌张力（tonin）的能力而得名。5-羟色胺的生物学效应被认为是通过刺激质膜上的5-羟色胺受体来实现的。5-羟色胺最近被证明是在血管平滑肌中合成的，并从外部来源吸收，将5-羟色胺置于细胞内。谷氨酰胺转胺酶利用伯胺如5-HT共价修饰谷氨酰胺残基上的蛋白质。我们验证了5-羟色胺是动脉血管平滑肌中转谷氨酰胺酶的底物的假设，并与蛋白质血清素化具有生理功能。以大鼠主动脉和培养的主动脉平滑肌细胞为模型。Western分析表明，转谷氨酰胺酶II存在于血管组织中，并观察到转谷氨酰胺酶的活性，因为它可以抑制游离胺戊胺生物素进入动脉蛋白。通过串联质谱分析，血清素生物素被掺入α -肌动蛋白、β-肌动蛋白、γ-肌动蛋白、肌球蛋白重链和丝蛋白A中。使用针对生物素或5-羟色胺的抗体，免疫沉淀和免疫细胞化学证实了平滑肌α -肌动蛋白的血清素化。重要的是，α-肌动蛋白依赖的动脉等距收缩到5-HT的过程被半胺减少。5-HT共价修饰与收缩性和细胞骨架相关的蛋白质。这些发现提示了5-羟色胺在血管平滑肌中的新的作用机制，并考虑了原胺在细胞内的作用。
Serotonin (5-hydroxytryptamine, 5-HT) was named for its source (sero-) and ability to modify smooth muscle tone (tonin). The biological effects of 5-HT are believed to be carried out by stimulation of serotonin receptors at the plasma membrane. Serotonin has recently been shown to be synthesized in vascular smooth muscle and taken up from external sources, placing 5-HT inside the cell. The enzyme transglutaminase uses primary amines such as 5-HT to covalently modify proteins on glutamine residues. We tested the hypothesis that 5-HT is a substrate for transglutaminase in arterial vascular smooth muscle, with protein serotonylation having physiological function. The model was the rat aorta and cultured aortic smooth muscle cells. Western analysis demonstrated that transglutaminase II was present in vascular tissue, and transglutaminase activity was observed as a cystamine-inhibitable incorporation of the free amine pentylamine-biotin into arterial proteins. Serotonin-biotin was incorporated into α -actin, β-actin, γ-actin, myosin heavy chain and filamin A as shown through tandem mass spectrometry. Using antibodies directed against biotin or 5-HT, immunoprecipitation and immunocytochemistry confirmed serotonylation of smooth muscle α–actin. Importantly, the α-actin-dependent process of arterial isometric contraction to 5-HT was reduced by cystamine. 5-HT covalently modifies proteins integral to contractility and the cytoskeleton. These findings suggest new mechanisms of action for 5-HT in vascular smooth muscle and consideration for intracellular effects of primary amines.
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影响因子：
1.7
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