Fleminger G, Howells R D, Kilpatrick D L, Udenfriend S
Proc Natl Acad Sci U S A. 1984 Dec;81(24):7985-8. doi: 10.1073/pnas.81.24.7985.
We have shown previously that enkephalin-containing peptides of high molecular weight increase 10- to 15-fold 3 days after adrenal denervation and have suggested that much of this material may represent newly synthesized proenkephalin. In this paper we report that most of the material appearing after denervation is indeed intact proenkephalin. The putative proenkephalin was partially purified by gel filtration chromatography and HPLC. A molecular size of about 25 kDa was estimated by gel filtration. On HPLC, the putative proenkephalin was eluted at a much higher propanol concentration than the 18.2-kDa enkephalin-containing peptide isolated previously. Treatment of this putative proenkephalin with endoproteinase Lys-C showed it contain [Met]-enkephalin, [Leu]enkephalin, [Met]enkephalin-Arg6-Phe7, and [Met]enkephalin-Arg6-Gly7-Leu8 in the same ratios as are found in proenkephalin as deduced from sequencing of proenkephalin cDNA.
我们之前已经表明,肾上腺去神经支配3天后,高分子量含脑啡肽的肽增加了10到15倍,并且我们认为这种物质的大部分可能代表新合成的脑啡肽原。在本文中,我们报告去神经支配后出现的大部分物质确实是完整的脑啡肽原。假定的脑啡肽原通过凝胶过滤色谱法和高效液相色谱法进行了部分纯化。通过凝胶过滤估计分子大小约为25 kDa。在高效液相色谱上,假定的脑啡肽原在比先前分离的18.2 kDa含脑啡肽的肽更高的丙醇浓度下洗脱。用内肽酶Lys-C处理这种假定的脑啡肽原表明它含有[Met]-脑啡肽、[Leu]脑啡肽、[Met]脑啡肽-Arg6-Phe7和[Met]脑啡肽-Arg6-Gly7-Leu8,其比例与从脑啡肽原cDNA测序推导的脑啡肽原中发现的比例相同。