Back to Search Start Over

Small molecule microarray screening methodology based on surface plasmon resonance imaging

Authors :
Vikramjeet Singh
Kuldeep Singh
Amita Nand
Huanqin Dai
Jianguo Wang
Lixin Zhang
Alejandro Merino
Jingsong Zhu
Source :
Arabian Journal of Chemistry, Vol 12, Iss 8, Pp 2111-2117 (2019)
Publication Year :
2019
Publisher :
Elsevier, 2019.

Abstract

In order to increase the scope and utility of small molecule microarrays (SMMs) we have combined SMMs and SPRi to screen small molecule antagonists against protein targets. Several small molecules, including immunosuppressive drugs (rapamycin and FK506) and reported inhibitors (FOBISIN and Blapsin) of 14-3-3ζ proteins have been used to validate this technology. Furthermore, a small library of isatin derivatives have been synthesized and screened on developed platform against 14-3-3ζ protein. Three molecules, derived from the endogenous intermediate isatin termed, FZIB-35, FZIB-36 and FZIB-38 were identified as novel inhibitors which shows significant interaction with 14-3-3ζ. A mutation in the binding groove of 14-3-3ζ, (K49E), almost abolishes the binding of these compounds to 14-3-3ζ protein. To exclude the probability of false positives, two more purified proteins (PtpA and BirA) were also tested. Furthermore, in order to confirm the binding pocket specificity, competition assay against R18 peptide was also carried out on presented platform. We show that SMMs in combination with SPRi are a powerful method to identify lead compounds in high throughput manner without the need to develop an activity based assay. Keywords: Small molecule microarray, Surface plasmon resonance, 14-3-3ζ protein, Isatin and ligand–protein interaction

Subjects

Subjects :
Chemistry
QD1-999

Details

Language :
English
ISSN :
18785352
Volume :
12
Issue :
8
Database :
Directory of Open Access Journals
Journal :
Arabian Journal of Chemistry
Publication Type :
Academic Journal
Accession number :
edsdoj.11d8018c4274ecf9288ad2e6fd79a20
Document Type :
article
Full Text :
https://doi.org/10.1016/j.arabjc.2014.12.020