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題名 | Identification of Subcellular Location of HCN4 Channel with D553N Mutant=具D553N突變之HCN4通道蛋白於亞細胞定位之辨識 |
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作者姓名(中文) | 李貫綸; 林彥昌; | 書刊名 | 華岡農科學報 |
卷期 | 37 2016.06[民105.06] |
頁次 | 頁1-8 |
分類號 | 368.5 |
關鍵詞 | 共定位; D553N; HCN4; Colocalization; |
語文 | 英文(English) |
中文摘要 | D553N是在HCN4通道蛋白C端的錯義突變(missense mutation)。在患者身上發現會導致包括長QT綜合徵、室性心動過速的多態性的嚴重心律失常,且會誘發竇房結功能障礙性心動過緩。雖然已經證實是由於它的轉運缺陷,造成的這種因突變導致的心律失常,並沒有分析顯示其於亞細胞(subcellular)水平的作用機制。根據本研究,我們證明了運輸缺陷突變HCN4 D553N僅會發生於內質網(endoplasmic reticulum, ER)和核週區域(perinuclear region)。轉染D553N與ER maker KKXX於HEK293細胞中兩天後,可以觀察到這兩種蛋白質的共定位(colocalization)。此外,我們所用的膜表面染劑FMI 34,不能與共定位,相反地,野生型(wild type)的HCN4通道能與細胞膜上的染劑FM1-34共定位,故將會以此做為對照。根據我們的實驗結果表明,該突變體D553N的亞細胞會保留在ER上,導致在細胞膜上該突變體喪失功能。 |
英文摘要 | D553N is a missense mutation in the C-linker of HCN4 channel protein. It was found in a patient with severe cardiac arrhythmias including long QT syndrome, ventricular tachycardia polymorphisms, and bradycardia induced by sinus node dysfunction. Although it has been implicated that the cardiac arrhythmia caused by this mutant was due to its trafficking defect, no analysis reveals the underlying mechanism in the subcellular level. Here, we demonstrated that the trafficking defective mutant HCN4 D553N is restricted in the endoplasmic reticulum (ER) and perinuclear region. Two days after transfection D553N with the ER maker KKXX in the HEK293 cells, colocalization of these two proteins could be observed. Further, we employed the membrane surface dye FM1-34 to show that D553N cannot colocalize with the membrane dye FM1-34. On the contrary, wild type HCN4 channels colocalized with the dye FM1-34 on the cell membrane which serves as the control for the comparison. In conclusion, our data further demonstrate the subcellular location that the D553N mutant is retained in the ER, resulting in loss-of-function of this mutant at the cell membrane. |
本系統之摘要資訊系依該期刊論文摘要之資訊為主。