Yoshizaki K, de Bock V, Solomon S
Life Sci. 1984 Feb 27;34(9):835-43. doi: 10.1016/0024-3205(84)90200-5.
Four different forms of bombesin-like immunoreactive peaks were detected in extracts of human fetal lung by the use of reversed-phase high performance liquid chromatography (HPLC). Peaks I, II, III and IV, (increasing retention time), were eluted using a 14-38% of acetonitrile gradient containing 0.1% trifluoroacetic acid (TFA). Peak II was the major material found in the extract of human fetal lung obtained at 16-20 weeks gestation. None of the four compounds contained in the eluted peaks had the same retention time as amphibian bombesin or porcine gastrin releasing peptide (GRP). On reversed-phase HPLC using two different solvent systems TFA or heptafluorobutyric acid (HFBA) as a hydrophobic counter ion, and in gel filtration chromatography, the chromatographic behavior of the main peak (peak II) was the same as that of the carboxyl terminal fragments of GRP, GRP18-27 or GRP19-27. This suggested that the peptide(s) in peak II resembled in composition the carboxy terminal 9 or 10 amino acids of porcine GRP. Following tryptic digestion the material in peak IV was converted to the more polar compound present in peak II. Two other peptide peaks were eluted close to peak II and these were presumed to be a modification of this main peak. One of the possible biosynthetic steps in the formation of bombesin-like peptides in human fetal lung could be a tryptic conversion of a less polar peptide to a more polar form (peak IV to II).
通过反相高效液相色谱法（HPLC）在人胎儿肺提取物中检测到四种不同形式的蛙皮素样免疫反应峰。峰I、II、III和IV（保留时间递增），使用含有0.1%三氟乙酸（TFA）的14 - 38%乙腈梯度洗脱。峰II是在妊娠16 - 20周时获得的人胎儿肺提取物中发现的主要物质。洗脱峰中所含的四种化合物均没有与两栖类蛙皮素或猪胃泌素释放肽（GRP）相同的保留时间。在使用两种不同溶剂系统（TFA或七氟丁酸（HFBA）作为疏水抗衡离子）的反相HPLC以及凝胶过滤色谱中，主峰（峰II）的色谱行为与GRP的羧基末端片段、GRP18 - 27或GRP19 - 27相同。这表明峰II中的肽在组成上类似于猪GRP的羧基末端9个或10个氨基酸。经胰蛋白酶消化后，峰IV中的物质转化为峰II中存在的极性更强的化合物。另外两个肽峰在峰II附近洗脱，推测它们是该主峰的修饰形式。人胎儿肺中蛙皮素样肽形成的可能生物合成步骤之一可能是一种极性较小的肽经胰蛋白酶转化为极性更强的形式（峰IV到峰II）。
