Clay K L, Murphy R C, Andres J L, Lynch J, Henson P M
Biochem Biophys Res Commun. 1984 Jun 29;121(3):815-25. doi: 10.1016/0006-291x(84)90751-4.
Platelet activating factor (PAF) synthesized by human neutrophils challenged by opsonized zymosan or calcium ionophore was isolated from cells and buffer using Bligh and Dyer extraction following the addition of tracer amounts of tritiated-PAF. The extract was subjected to TLC separation of phospholipid classes, followed by reverse phase HPLC for molecular species separation. All fractions were measured for radioactivity, biological activity and fast atom bombardment mass spectrometry. While the radioactive tracer PAF could be separated into three molecular species, PAF biological activity eluted as a single component which was characterized as 1-O-hexadecyl-2-acetyl-glycero-3-phosphocholine. The lack of molecular species heterogeneity of PAF produced in response to stimuli implies a higher degree of control of biosynthesis than previously suspected.
用调理酵母聚糖或钙离子载体刺激人中性粒细胞后合成的血小板活化因子（PAF），在加入微量氚标记的PAF后，采用布莱和戴尔萃取法从细胞和缓冲液中分离出来。提取物先进行磷脂类的薄层色谱分离，然后进行反相高效液相色谱以分离分子种类。对所有馏分进行放射性、生物活性和快原子轰击质谱测定。虽然放射性示踪剂PAF可分离为三种分子种类，但PAF生物活性以单一成分洗脱，其特征为1-O-十六烷基-2-乙酰基甘油-3-磷酸胆碱。对刺激产生的PAF缺乏分子种类异质性，这意味着生物合成的控制程度比以前认为的更高。
