Schmidt M F
EMBO J. 1984 Oct;3(10):2295-300. doi: 10.1002/j.1460-2075.1984.tb02129.x.
[3H]Myristic and [3H]palmitic acid were compared as tracers for the fatty acylation of cellular lipids and viral glycoproteins in chicken embryo cells infected with fowl plague and Semliki Forest virus (SFV). Both of these substrates are incorporated into glycerolipids to a similar extent, whereas sphingolipids show much higher levels of palmitate than myristate after a 20 h labeling period. Both fatty acid species were found to be subject to metabolic conversions into longer chain fatty acids yielding 11.7% C16:0 from [3H]myristic and 11.8% C18:0 from [3H]palmitic acid. The reverse, a metabolic shortening of the exogenous acyl-chains yielding, for instance, significant levels of myristic acid from palmitic acid was not observed. Out of the various [3H]fatty acids present after in vivo labeling with [3H]myristic acid (C14:0) the elongated acyl-species arising from metabolic conversion (e.g., C16:0; C18:0) are preferred over myristic acid in the acylation of SFV E1 and E2 and of the influenza viral hemagglutinin (HA2). During acylation of exogenous E1 from SFV in vitro incorporation of palmitic acid from palmitoyl CoA exceeds that of myristic acid from myristoyl CoA by a factor of 37. This indicates that specificity for the incorporation of fatty acids into viral membrane proteins occurs at the level of the polypeptide acyltransferase(s).
将[3H]肉豆蔻酸和[3H]棕榈酸作为示踪剂,用于研究感染禽瘟病毒和Semliki森林病毒(SFV)的鸡胚细胞中细胞脂质和病毒糖蛋白的脂肪酰化作用。这两种底物掺入甘油脂质的程度相似,而在标记20小时后,鞘脂中棕榈酸的含量远高于肉豆蔻酸。发现这两种脂肪酸都会代谢转化为长链脂肪酸,[3H]肉豆蔻酸产生11.7%的C16:0,[3H]棕榈酸产生11.8%的C18:0。未观察到相反的情况,即外源酰基链的代谢缩短,例如从棕榈酸产生大量肉豆蔻酸。在用[3H]肉豆蔻酸(C14:0)进行体内标记后存在的各种[3H]脂肪酸中,由代谢转化产生的延长酰基物种(例如C16:0;C18:0)在SFV E1和E2以及流感病毒血凝素(HA2)的酰化过程中比肉豆蔻酸更受青睐。在体外对SFV的外源E1进行酰化时,来自棕榈酰辅酶A的棕榈酸掺入量比来自肉豆蔻酰辅酶A的肉豆蔻酸高出37倍。这表明脂肪酸掺入病毒膜蛋白的特异性发生在多肽酰基转移酶水平。