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Identification and validation of VEGFR2 kinase as a target of voacangine by a systematic combination of DARTS and MSI

Kim, Yonghyo LU ; Sugihara, Yutaka LU ; Kim, Tae Young ; Cho, Sung Min ; Kim, Jin Young ; Lee, Ju Yeon ; Yoo, Jong Shin ; Song, Doona ; Han, Gyoonhee and Rezeli, Melinda LU , et al. (2020) In Biomolecules 10(4).
Abstract

Although natural products are an important source of drugs and drug leads, identification and validation of their target proteins have proven difficult. Here, we report the development of a systematic strategy for target identification and validation employing drug affinity responsive target stability (DARTS) and mass spectrometry imaging (MSI) without modifying or labeling natural compounds. Through a validation step using curcumin, which targets aminopeptidase N (APN), we successfully standardized the systematic strategy. Using label-free voacangine, an antiangiogenic alkaloid molecule as the model natural compound, DARTS analysis revealed vascular endothelial growth factor receptor 2 (VEGFR2) as a target protein. Voacangine inhibits... (More)

Although natural products are an important source of drugs and drug leads, identification and validation of their target proteins have proven difficult. Here, we report the development of a systematic strategy for target identification and validation employing drug affinity responsive target stability (DARTS) and mass spectrometry imaging (MSI) without modifying or labeling natural compounds. Through a validation step using curcumin, which targets aminopeptidase N (APN), we successfully standardized the systematic strategy. Using label-free voacangine, an antiangiogenic alkaloid molecule as the model natural compound, DARTS analysis revealed vascular endothelial growth factor receptor 2 (VEGFR2) as a target protein. Voacangine inhibits VEGFR2 kinase activity and its downstream signaling by binding to the kinase domain of VEGFR2, as was revealed by docking simulation. Through cell culture assays, voacangine was found to inhibit the growth of glioblastoma cells expressing high levels of VEGFR2. Specific localization of voacangine to tumor compartments in a glioblastoma xenograft mouse was revealed by MSI analysis. The overlap of histological images with the MSI signals for voacangine was intense in the tumor regions and showed colocalization of voacangine and VEGFR2 in the tumor tissues by immunofluorescence analysis of VEGFR2. The strategy employing DARTS and MSI to identify and validate the targets of a natural compound as demonstrated for voacangine in this study is expected to streamline the general approach of drug discovery and validation using other biomolecules including natural products.

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organization
publishing date
type
Contribution to journal
publication status
published
subject
keywords
Anti-angiogenesis, Curcumin, Label-free method for drugs, Mechanism of action, Natural products, Receptor tyrosine kinases, Target identification, Target validation
in
Biomolecules
volume
10
issue
4
article number
508
publisher
Multidisciplinary Digital Publishing Institute (MDPI)
external identifiers
  • pmid:32230857
  • scopus:85082730126
ISSN
2218-273X
DOI
10.3390/biom10040508
language
English
LU publication?
yes
id
c03e9a17-287b-41a0-9632-9e3bbff40c49
date added to LUP
2020-04-16 17:36:15
date last changed
2021-06-23 05:06:23
@article{c03e9a17-287b-41a0-9632-9e3bbff40c49,
  abstract     = {<p>Although natural products are an important source of drugs and drug leads, identification and validation of their target proteins have proven difficult. Here, we report the development of a systematic strategy for target identification and validation employing drug affinity responsive target stability (DARTS) and mass spectrometry imaging (MSI) without modifying or labeling natural compounds. Through a validation step using curcumin, which targets aminopeptidase N (APN), we successfully standardized the systematic strategy. Using label-free voacangine, an antiangiogenic alkaloid molecule as the model natural compound, DARTS analysis revealed vascular endothelial growth factor receptor 2 (VEGFR2) as a target protein. Voacangine inhibits VEGFR2 kinase activity and its downstream signaling by binding to the kinase domain of VEGFR2, as was revealed by docking simulation. Through cell culture assays, voacangine was found to inhibit the growth of glioblastoma cells expressing high levels of VEGFR2. Specific localization of voacangine to tumor compartments in a glioblastoma xenograft mouse was revealed by MSI analysis. The overlap of histological images with the MSI signals for voacangine was intense in the tumor regions and showed colocalization of voacangine and VEGFR2 in the tumor tissues by immunofluorescence analysis of VEGFR2. The strategy employing DARTS and MSI to identify and validate the targets of a natural compound as demonstrated for voacangine in this study is expected to streamline the general approach of drug discovery and validation using other biomolecules including natural products.</p>},
  author       = {Kim, Yonghyo and Sugihara, Yutaka and Kim, Tae Young and Cho, Sung Min and Kim, Jin Young and Lee, Ju Yeon and Yoo, Jong Shin and Song, Doona and Han, Gyoonhee and Rezeli, Melinda and Welinder, Charlotte and Appelqvist, Roger and Marko-Varga, György and Kwon, Ho Jeong},
  issn         = {2218-273X},
  language     = {eng},
  month        = {03},
  number       = {4},
  publisher    = {Multidisciplinary Digital Publishing Institute (MDPI)},
  series       = {Biomolecules},
  title        = {Identification and validation of VEGFR2 kinase as a target of voacangine by a systematic combination of DARTS and MSI},
  url          = {http://dx.doi.org/10.3390/biom10040508},
  doi          = {10.3390/biom10040508},
  volume       = {10},
  year         = {2020},
}