Characterization of mutant serine palmitoyltransferase 1 in LY-B cells.
Characterization of mutant serine palmitoyltransferase 1 in LY-B cells.
复制标题
LY-B 细胞中突变丝氨酸棕榈酰转移酶 1 的表征。
DOI：
10.1007/s11745-009-3316-4
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发表时间：
2009
期刊：
影响因子：
1.9
通讯作者：
Hanada,Kentaro
中科院分区：
文献类型：
--
作者：
Momin,AminA;Park,Hyejung;Allegood,JeremyC;Leipelt,Martina;Kelly,SamuelL;MerrillJr,AlfredH;Hanada,Kentaro
CHO-LY-B细胞由于缺乏丝氨酸棕榈酰转移酶（SPT）活性而被用于鞘脂代谢和功能的研究，LY-B细胞的SPT 1转录物的克隆和测序鉴定为核苷酸738处鸟嘌呤至腺嘌呤的突变，引起G246 R转化。蛋白质印迹显示低表达的突变体SPT 1肽，但活性是不可检测的质谱分析[13 C]-棕榈酸掺入二氢鞘氨醇，鞘氨醇，1-脱氧二氢鞘氨醇，或1-脱氧甲基二氢鞘氨醇。用化学伴侣（DMSO或甘油）处理LY-B细胞增加了突变体SPT 1和SPT 2的数量，但SPT活性没有恢复。本研究已经确定仓鼠SPT 1中的G246 R突变导致SPT活性丧失。
CHO-LY-B cells have been useful in studies of sphingolipid metabolism and function because they lack serine palmitoyltransferase (SPT) activity.Cloning and sequencing of the SPT1 transcript of LY-B cells identified the mutation as a guanine to adenine change at nucleotide 738, causing a G246R transformation. Western blots revealed low expression of the mutant SPT1 peptide, but activity was not detectable by mass spectrometric analysis of [13C]-palmitate incorporation into sphinganine, sphingosine, 1-deoxysphinganine, or 1-desoxymethylsphinganine. Treatment of LY-B cells with chemical chaperones (DMSO or glycerol) increased the amounts of mutant SPT1 as well as SPT2, but SPT activity was not restored. This study has established that G246R mutation in hamster SPT1 results in the loss of SPT activity.