Lipid-modified, cysteinyl-containing peptides of diverse structures are efficiently S-acylated at the plasma membrane of mammalian cells.
Lipid-modified, cysteinyl-containing peptides of diverse structures are efficiently S-acylated at the plasma membrane of mammalian cells.
复制标题
在哺乳动物细胞的质膜上有效地将各种结构的脂质修饰的含有胱烷基的肽有效地覆盖。
DOI：
10.1083/jcb.134.3.647
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发表时间：
1996-08
影响因子：
7.8
通讯作者：
Silvius, J R
中科院分区：
文献类型：
--
作者：
Schroeder, H;Leventis, R;Shahinian, S;Walton, P A;Silvius, J R
发现多种含半胱氨酸的脂质修饰的肽被培养的哺乳动物细胞S-酰化。酰化反应对半胱氨酰的特异性高于对丝氨酸残基的特异性，对脂质修饰的肽的特异性高于对亲水性肽的特异性。根据各种标准，S-酰化过程似乎是酶促的，并且在各种特征上类似于质膜相关蛋白的S-酰化，包括被衣霉素抑制。S-酰化反应的底物范围包括但不限于掺入肉豆蔻酰GC-和-CXC（法呢基）-OCH 3基序的脂肽，其在各种细胞内蛋白中可逆地S-酰化。质谱分析表明，棕榈酰残基构成了与携带肉豆蔻酰GC-基序的脂肽偶联的S-酰基的主要但不是唯一类型，也可检测到少量的S-硬脂酰和S-油酰取代基。使用NBD标记的半胱氨酰脂肽的荧光显微镜显示，脂肽S-酰化的产物，它不能在膜之间扩散，在几乎所有的情况下，优先定位于质膜。即使在降低的温度下也发现这种优先定位，其中从高尔基体复合体到质膜的囊泡运输被抑制，强烈表明质膜本身是这些物种的S-酰化的优选位点。独特的脂肽研究，物种纳入非生理N-肉豆蔻酰半胱氨酰基序也显示大量形成的S-酰化产物在第二，细胞内隔室确定为高尔基复合体，其标记与荧光神经酰胺。我们的研究结果表明，不同的S-酰基转移酶存在于高尔基体复合体和质膜隔室和S-酰化的图案，如肉豆蔻酰GC-发生在质膜特异性，提供有效的靶向细胞蛋白轴承这种图案的膜隔室。
A variety of cysteine-containing, lipid-modified peptides are found to be S-acylated by cultured mammalian cells. The acylation reaction is highly specific for cysteinyl over serinyl residues and for lipid- modified peptides over hydrophilic peptides. The S-acylation process appears by various criteria to be enzymatic and resembles the S- acylation of plasma membrane-associated proteins in various characteristics, including inhibition by tunicamycin. The substrate range of the S-acylation reaction encompasses, but is not limited to, lipopeptides incorporating the motifs myristoylGC- and -CXC(farnesyl)- OCH3, which are reversibly S-acylated in various intracellular proteins. Mass-spectrometric analysis indicates that palmitoyl residues constitute the predominant but not the only type of S-acyl group coupled to a lipopeptide carrying the myristoylGC- motif, with smaller amounts of S-stearoyl and S-oleoyl substituents also detectable. Fluorescence microscopy using NBD-labeled cysteinyl lipopeptides reveals that the products of lipopeptide S-acylation, which cannot diffuse between membranes, are in almost all cases localized preferentially to the plasma membrane. This preferential localization is found even at reduced temperatures where vesicular transport from the Golgi complex to the plasma membrane is suppressed, strongly suggesting that the plasma membrane itself is the preferred site of S- acylation of these species. Uniquely among the lipopeptides studied, species incorporating an unphysiological N-myristoylcysteinyl- motif also show substantial formation of S-acylated products in a second, intracellular compartment identified as the Golgi complex by its labeling with a fluorescent ceramide. Our results suggest that distinct S-acyltransferases exist in the Golgi complex and plasma membrane compartments and that S-acylation of motifs such as myristoylGC- occurs specifically at the plasma membrane, affording efficient targeting of cellular proteins bearing such motifs to this membrane compartment.