Roth K A, Evans C J, Lorenz R G, Weber E, Barchas J D, Chang J K
Biochem Biophys Res Commun. 1983 Apr 29;112(2):528-36. doi: 10.1016/0006-291x(83)91497-3.
Rat and guinea pig brain extracts were examined for the occurrence of gastrin-releasing peptide (GRP)-like substances by sequence specific radioimmunoassays interfaced with gel filtration and reversed phase high performance liquid chromatography (RP-HPLC). Tryptic digestion of the immunoreactive peptides followed by RP-HPLC was used to further characterize GRP-related peptides in brain. Using these analytical techniques it was found that guinea pig brain extracts contained a peptide with characteristics identical to authentic GRP (27 amino acid residues long). A carboxyterminal fragment with the characteristics of GRP(18-27) as well as a respective aminoterminal fragment with the characteristics of GRP(1-16) were also present in guinea pig brain extracts. The GRP(18-27) seems to correspond to the bombes in related material that has been described previously in mammalian brain extracts. Rat brain extracts also contained a peptide with the characteristics of GRP(18-27). The corresponding aminoterminal fragment, however, behaved differently on RP-HPLC from authentic GRP(1-16) and it was not recognized by antibodies directed to the aminoterminal tridecapeptide fragment of authentic GRP. Similarly the GRP-like peptide from rat brain did not comigrate on RP-HPLC with authentic GRP and was unreactive to antibodies directed toward the aminoterminus of GRP.
通过与凝胶过滤和反相高效液相色谱（RP-HPLC）联用的序列特异性放射免疫分析法，检测大鼠和豚鼠脑提取物中胃泌素释放肽（GRP）样物质的存在情况。对免疫反应性肽进行胰蛋白酶消化，然后进行RP-HPLC，以进一步鉴定脑中与GRP相关的肽。使用这些分析技术发现，豚鼠脑提取物中含有一种肽，其特性与天然GRP（27个氨基酸残基）相同。豚鼠脑提取物中还存在具有GRP（18-27）特性的羧基末端片段以及具有GRP（1-16）特性的相应氨基末端片段。GRP（18-27）似乎与先前在哺乳动物脑提取物中描述的相关物质中的铃蟾肽相对应。大鼠脑提取物中也含有一种具有GRP（18-27）特性的肽。然而，相应的氨基末端片段在RP-HPLC上的行为与天然GRP（1-16）不同，并且不被针对天然GRP氨基末端十三肽片段的抗体识别。同样，来自大鼠脑的GRP样肽在RP-HPLC上与天然GRP不共迁移，并且对针对GRP氨基末端的抗体无反应。
