Krueger J M, Pappenheimer J R, Karnovsky M L
Proc Natl Acad Sci U S A. 1978 Oct;75(10):5235-8. doi: 10.1073/pnas.75.10.5235.
Sleep-promoting factor was purified from acid/acetone extracts of whole brains of rabbits and from brainstems of slaughterhouse cattle. Intraventricular infusion of extracts purified by means of ion exchange and gel filtration induced excess slow-wave sleep in rabbits for 5-10 hr. The procedure is simple and provides material suitable for physiological studies. Further treatment by partition chromatography and electrophoresis yielded an active product that was purified at least 1 million-fold. This product was inactivated by incubation with mixed carboxypeptidases A and B. Amino acid analysis of acid hydrolysates indicated that the effective dose was less than 150 pmol per rabbit and the original concentration in brain tissue was of the order of 30 pmol/g of brain.
促睡眠因子是从兔全脑的酸/丙酮提取物以及屠宰场牛的脑干中纯化得到的。通过离子交换和凝胶过滤纯化后的提取物脑室内注射可使兔产生5至10小时的过度慢波睡眠。该方法简单，且提供了适合生理学研究的材料。通过分配色谱法和电泳进一步处理得到了一种活性产物，其纯化倍数至少为100万倍。该产物与羧肽酶A和B混合孵育后失活。酸水解产物的氨基酸分析表明，每只兔的有效剂量小于150皮摩尔，脑组织中的原始浓度约为30皮摩尔/克脑。
