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Conjugates of adenosine mimetics and arginine-rich peptides serve as inhibitors and fluorescent probes but not as long-lifetime photoluminescent probes for protein arginine methyltransferases
Journal of Peptide Science ( IF 2.1 ) Pub Date : 2022-10-08 , DOI: 10.1002/psc.3456
Darja Lavogina 1, 2 , Naila Nasirova 1 , Tanel Sõrmus 1 , Taavo Tähtjärv 1 , Erki Enkvist 1 , Kaido Viht 1 , Tõiv Haljasorg 1 , Koit Herodes 1 , Jana Jaal 2, 3 , Asko Uri 1
Affiliation  

The conjugates of an adenosine mimetic and oligo-l-arginine or oligo-d-arginine (ARCs) were initially designed in our research group as inhibitors and photoluminescent probes targeting basophilic protein kinases. Here, we explored a panel of ARCs and their fluorescent derivatives in biochemical assays with members of the protein arginine methyltransferase (PRMT) family, focusing specifically on PRMT1. In the binding/displacement assay with detection of fluorescence anisotropy, we found that ARCs and arginine-rich peptides could serve as high-affinity ligands for PRMT1, whereas the equilibrium dissociation constant values depended dramatically on the number of arginine residues within the compounds. The fluorescently labeled probe ARC-1081 was displaced from its complex with PRMT1 by both S-adenosyl-l-methionine (SAM) and S-adenosyl-l-homocysteine (SAH), indicating binding of the adenosine mimetic of ARCs to the SAM/SAH-binding site within PRMT1. The ARCs that had previously shown microsecond-lifetime photoluminescence in complex with protein kinases did not feature such property in complex with PRMT1, demonstrating the selectivity of the time-resolved readout format. When tested against a panel of PRMT family members in single-dose inhibition experiments, a micromolar concentration of ARC-902 was required for the inhibition of PRMT1 and PRMT7. Overall, our results suggest that the compounds containing multiple arginine residues (including the well-known cell-penetrating peptides) are likely to inhibit PRMT and thus interfere with the epigenetic modification status in complex biological systems, which should be taken into consideration during interpretation of the experimental data.

中文翻译:

腺苷模拟物和富含精氨酸的肽的结合物用作抑制剂和荧光探针,但不能用作蛋白质精氨酸甲基转移酶的长寿命光致发光探针

腺苷模拟物和寡-l-精氨酸或寡-d的结合物-精氨酸 (ARC) 最初是在我们的研究小组中设计的,作为针对嗜碱性蛋白激酶的抑制剂和光致发光探针。在这里,我们与蛋白质精氨酸甲基转移酶 (PRMT) 家族的成员一起探索了一组 ARC 及其在生化测定中的荧光衍生物,特别关注 PRMT1。在检测荧光各向异性的结合/置换测定中,我们发现 ARC 和富含精氨酸的肽可以作为 PRMT1 的高亲和力配体,而平衡解离常数值显着取决于化合物中精氨酸残基的数量。荧光标记的探针 ARC-1081 被S-腺苷-l-甲硫氨酸 (SAM) 和S-腺苷- l-同型半胱氨酸 (SAH),表明 ARC 的腺苷模拟物与 PRMT1 内的 SAM/SAH 结合位点结合。先前在与蛋白激酶复合物中显示微秒寿命光致发光的 ARC 在与 PRMT1 复合物中不具有这种特性,证明了时间分辨读出格式的选择性。当在单剂量抑制实验中针对一组 PRMT 家族成员进行测试时,需要微摩尔浓度的 ARC-902 来抑制 PRMT1 和 PRMT7。总的来说,我们的结果表明,含有多个精氨酸残基的化合物(包括众所周知的细胞穿透肽)可能会抑制 PRMT,从而干扰复杂生物系统中的表观遗传修饰状态,
更新日期:2022-10-08
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