LC/MS evaluation of metabolism and membrane transport of bombesin peptides.
LC/MS evaluation of metabolism and membrane transport of bombesin peptides.
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DOI：
10.1007/s00726-010-0696-y
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发表时间：
2011-02
期刊：
影响因子：
3.5
通讯作者：
Chen, Xiaoyuan
中科院分区：
文献类型：
--
作者：
Gu, Dongyu;Ma, Ying;Niu, Gang;Yan, Yongjun;Lang, Lixin;Aisaand, Haji Akber;Gao, Haokao;Kiesewetter, Dale O.;Chen, Xiaoyuan
评价了两种bombsin肽，GRPR激动剂[fQWAVGHL-NH 2]和拮抗剂[fQWAVGHL-NH乙基]。我们采用高灵敏度的沃茨Q-Tof Premier MS结合UPLC系统鉴定大鼠肝细胞或PC-3人前列腺癌细胞产生的代谢产物;采用具有高灵敏度定量和定性性能的AB/MDS 4000 Q-Trap LC/MS/MS系统定量分析GRPR激动剂和拮抗剂在PC-3细胞中的内化。GRPR激动剂和拮抗剂的主要代谢产物均为W和A之间的肽键水解的结果，激动剂的N-末端片段m/z 446（fQW-OH）和拮抗剂的m/z 480（fQW-OH）的观察证实了这一点。两种肽也在A和V之间水解，对于激动剂形成峰m/z 517 [fQWA-OH]和m/z 555（VGHLM-NH 2），对于拮抗剂形成峰m/z 551 [fQWA-OH]和m/z 452（VGHL-NH乙基）。肽激动剂还形成了一种独特的代谢产物，这是由C-末端酰胺水解引起的。在大鼠肝细胞和PC-3细胞中，与激动剂相比，拮抗剂的代谢明显较慢。拮抗剂也显示出显著低于PC-3细胞内化率比激动剂。总之，通过LC/MS鉴定了GRPR激动剂和拮抗剂肽的代谢特征，在大鼠肝细胞孵育中，拮抗剂肽比激动剂肽更稳定。一个主要因素可能是与激动剂的未取代酰胺相比，C-末端L-NH乙基基团具有耐水解性。另一个因素可能是激动剂和拮抗剂的不同氨基酸序列，其也可能影响酶水解。拮抗剂配体可能更适用于受体靶向成像，主要是由于其较高的代谢稳定性。
Two bombsin peptides, GRPR agonist [Aca-QWAVGHLM-NH2] and antagonist [fQWAVGHL-NHEthyl] were evaluated. We employed the highly sensitive Waters Q-Tof Premier MS coupled with a UPLC system to identify the metabolites produced by rat hepatocytes or PC-3 human prostate cancer cells; and we utilized the AB/MDS 4000 Q-Trap LC/MS/MS system with highly sensitive quantitative and qualitative performance, to quantitatively analyze the internalization of GRPR agonist and antagonist in PC-3 cells. The major metabolites of both GRPR agonist and antagonist were the result of peptide bond hydrolysis between W and A which was demonstrated by observation of the N-terminal fragment m/z 446 (Aca-QW-OH) for agonist and m/z 480 (fQW-OH) for antagonist. Both peptides were also hydrolyzed between A and V which formed peaks m/z 517 [Aca-QWA-OH] and m/z 555 (VGHLM-NH2) for the agonist and m/z 551 [fQWA-OH] and m/z 452 (VGHL-NHEthyl) for the antagonist. The peptide agonist also formed a unique metabolite that resulted from hydrolysis of the C-terminal amide. The antagonist showed significantly slower metabolism as compared to the agonist in both rat hepatocytes and PC-3 cells. The antagonist also showed significantly lower PC-3 cell internalization rate than that of the agonist. In conclusion, the metabolism profiles of both GRPR agonist and antagonist peptides were identified by LC/MS. The antagonist peptide was more stable than the agonist peptide in rat hepatocyte incubation. One major factor could be the hydrolysis-resistant C-terminal L-NHEthyl group compared with the unsubstituted amide of the agonist. Another factor could be different amino acid sequences of the agonist and antagonist that may also influence the enzymatic hydrolysis. The antagonist ligand is potentially more useful for receptor-targeted imaging due primarily to its higher metabolic stability.
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DOI：
10.1007/s00259-002-1040-x
发表时间：
2003-02-01
影响因子：
9.1
作者：
Nock, B;Nikolopoulou, A;Maina, T
通讯作者：
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影响因子：
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作者：
Smith, CJ;Volkert, WA;Hoffman, TJ
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