Leblanc P, Capone J, Gerber G E
J Biol Chem. 1982 Dec 25;257(24):14586-9.
A synthesis of a homologous series of highly radioactive photoreactive fatty acids has been developed. Oxidative cleavage of the cis double bonds of unsaturated fatty acids by NaIO4/OsO4 produced the omega-oxo fatty acids; subsequent reduction by [3H]NaBH4 afforded a series of highly radioactive [omega-3H]hydroxy fatty acids to which a variety of photoreactive probes can be readily attached. In this study, the m-diazirinophenoxy group was coupled to the omega-hydroxy groups of the fatty acids, yielding the omega-(m-diazirinophenoxy) derivatives of nonanoic, undecanoic, tridecanoic, and pentadecanoic acid. These photoreactive derivatives were incorporated by L-cells into their major phospholipids; in addition, a number of proteins were shown to be acylated by these fatty acids. The photoreactive group was shown to remain intact as indicated by the cross-linking of the resulting photoreactive phospholipids to other phospholipids as well as to a number of L-cell proteins upon photolysis. This procedure thus identifies integral membrane proteins and directly produces photoaffinity derivatives of those proteins normally acylated by fatty acids.
已开发出一系列高放射性光反应性脂肪酸的合成方法。通过NaIO4/OsO4对不饱和脂肪酸的顺式双键进行氧化裂解，生成ω-氧代脂肪酸；随后用[3H]NaBH4还原，得到一系列高放射性的[ω-3H]羟基脂肪酸，各种光反应性探针可很容易地连接到这些脂肪酸上。在本研究中，间重氮苯氧基与脂肪酸的ω-羟基偶联，生成壬酸、十一烷酸、十三烷酸和十五烷酸的ω-(间重氮苯氧基)衍生物。这些光反应性衍生物被L细胞掺入其主要磷脂中；此外，一些蛋白质被证明可被这些脂肪酸酰化。光解后，所得光反应性磷脂与其他磷脂以及一些L细胞蛋白质发生交联，表明光反应性基团保持完整。因此，该方法可鉴定完整膜蛋白，并直接产生通常被脂肪酸酰化的那些蛋白的光亲和衍生物。
