Preparation and Application of a Chemical Probe for Identifying the Targets of the Marine Cyclic Peptide Kapakahine A.

Preparation and Application of a Chemical Probe for Identifying the Targets of the Marine Cyclic Peptide Kapakahine A.
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DOI:
10.3390/molecules27031072
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发表时间:
2022-02-05
期刊:
Molecules (Basel, Switzerland)
影响因子:
--
通讯作者:
Nakao Y
Nakao Y
中科院分区:
其他
文献类型:
--
作者:
Kamihira R;Nakao Y

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海洋生物是生物活性次级代谢产物的丰富来源。虽然许多具有生物活性的海洋天然产物已被分离,但成功阐明其作用机制仍然有限。在这项研究中,我们制备了一种基于海洋环肽kapakahine A(1)的探针分子,通过引入一个带有叠氮末端基团的接头,这使得能够引入荧光基团以有效监测亚细胞定位,或偶联到亲和珠以下拉靶蛋白。LC/MS/MS测量、ProteinPilot分析和Western印迹的结果表明,kapakahine A与线粒体内膜蛋白PHB 1、PHB 2和ANT 2相互作用,这与使用荧光探针的亚细胞定位分析的结果一致。
Marine organisms are a rich source of bioactive secondary metabolites. Although many marine natural products with bioactivities have been isolated, successful elucidation of their mechanisms of action remains limited. In this study, we prepared a probe molecule based on the marine cyclic peptide kapakahine A (1) by introducing a linker with an azide terminal group, which enables the introduction of fluorescent groups for the effective monitoring of subcellular localization, or coupling to affinity beads for the pull-down of target proteins. The results of LC/MS/MS measurements, ProteinPilot analysis, and Western blotting suggest that kapakahine A interacts with the mitochondrial inner membrane proteins PHB1, PHB2, and ANT2, which is consistent with the results of the subcellular localization analysis using a fluorescent probe.
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