An Integrative Biology Approach to Quantify the Biodistribution of Azidohomoalanine In Vivo
An Integrative Biology Approach to Quantify the Biodistribution of Azidohomoalanine In Vivo
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DOI：
10.1007/s12195-023-00760-4
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发表时间：
2023-03-23
影响因子：
2.8
通讯作者：
Kinzer-Ursem，Tamara L.
中科院分区：
文献类型：
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作者：
Saleh，Aya M.;VanDyk，Tyler G.;Kinzer-Ursem，Tamara L.
鉴定和定量新合成的蛋白质(NSP)对于了解蛋白质在发育和疾病中的动态变化至关重要。探测新生蛋白质组可以使用非典型氨基酸(NCAA)，利用内源翻译机制选择性地标记NSP，然后用质谱仪进行定量。我们先前已经证明，通过注射叠氮高丙氨酸(AHA)，一种NCAA和蛋氨酸(Met)的类似物，标记间尿液蛋白质组是可行的，而不需要Met耗尽。AHA标记可以解决生物学问题，其中时间蛋白质动力学是重要的。然而，要获得这种时间分辨率，需要更完整地了解AHA在组织中的分布动力学。结果为了解决这些差距，我们创建了一个确定的、隔室模型，用于研究AHA在小鼠体内的动力学转运和掺入。模型结果表明，能够预测AHA在各种组织和剂量范例中的分布和蛋白质标记。为了确定该方法在活体研究中的适用性，我们通过分析不同AHA给药方案后的血浆和肝脏代谢物，研究了AHA给药对正常生理的影响。结论我们的结果表明，我们可以重复性地预测蛋白质标记，并且在我们的实验研究过程中，给药这种类似物不会对生存生理学产生显著的改变。我们期待这个模型成为一个有用的工具来指导未来利用这一技术来研究蛋白质组对刺激的反应的实验。
BackgroundIdentification and quantitation of newly synthesized proteins (NSPs) are critical to understanding protein dynamics in development and disease. Probing the nascent proteome can be achieved using non-canonical amino acids (ncAAs) to selectively label the NSPs utilizing endogenous translation machinery, which can then be quantitated with mass spectrometry. We have previously demonstrated that labeling thein vivomurine proteome is feasible via injection of azidohomoalanine (Aha), an ncAA and methionine (Met) analog, without the need for Met depletion. Aha labeling can address biological questions wherein temporal protein dynamics are significant. However, accessing this temporal resolution requires a more complete understanding of Aha distribution kinetics in tissues.ResultsTo address these gaps, we created a deterministic, compartmental model of the kinetic transport and incorporation of Aha in mice. Model results demonstrate the ability to predict Aha distribution and protein labeling in a variety of tissues and dosing paradigms. To establish the suitability of the method forin vivostudies, we investigated the impact of Aha administration on normal physiology by analyzing plasma and liver metabolomes following various Aha dosing regimens. We show that Aha administration induces minimal metabolic alterations in mice.ConclusionsOur results demonstrate that we can reproducibly predict protein labeling and that the administration of this analog does not significantly alterin vivophysiology over the course of our experimental study. We expect this model to be a useful tool to guide future experiments utilizing this technique to study proteomic responses to stimuli.