scholarly journals Fluorescence imaging of drug target proteins using chemical probes

2020 ◽  
Vol 10 (5) ◽  
pp. 426-433 ◽  
Author(s):  
Hao Zhu ◽  
Itaru Hamachi
Keyword(s):  
Fluorescence Imaging ◽  
Drug Target ◽  
Chemical Probes ◽  
Target Proteins

scholarly journals Exploiting Chemical Probes for Identifying Drug Target Proteins

2003 ◽  
Vol 51 (5) ◽  
pp. 492-498 ◽  
Author(s):  
Toshitaka SATO ◽  
Yoshiya ODA ◽  
Takashi OWA ◽  
Takeshi NAGASU
Keyword(s):  
Drug Target ◽  
Chemical Probes ◽  
Target Proteins

Monitoring Drug Target Engagement in Cells and Tissues Using the Cellular Thermal Shift Assay

Science ◽  
2013 ◽  
Vol 341 (6141) ◽  
pp. 84-87 ◽  
Author(s):  
Daniel Martinez Molina ◽  
Rozbeh Jafari ◽  
Marina Ignatushchenko ◽  
Takahiro Seki ◽  
E. Andreas Larsson ◽  
...  
Keyword(s):  
Drug Target ◽  
Drug Transport ◽  
Drug Binding ◽  
Target Engagement ◽  
Tissue Samples ◽  
Target Proteins ◽  
Thermal Shift Assay ◽  
Cellular Thermal Shift Assay ◽  
Thermal Shift ◽  
In Cells

The efficacy of therapeutics is dependent on a drug binding to its cognate target. Optimization of target engagement by drugs in cells is often challenging, because drug binding cannot be monitored inside cells. We have developed a method for evaluating drug binding to target proteins in cells and tissue samples. This cellular thermal shift assay (CETSA) is based on the biophysical principle of ligand-induced thermal stabilization of target proteins. Using this assay, we validated drug binding for a set of important clinical targets and monitored processes of drug transport and activation, off-target effects and drug resistance in cancer cell lines, as well as drug distribution in tissues. CETSA is likely to become a valuable tool for the validation and optimization of drug target engagement.

Monitoring Drug-Target Interactions through Target Engagement-Mediated Amplification on Arrays and in situ

2021 ◽  
Author(s):  
Rasel Al-Amin ◽  
Lars Johansson ◽  
Eldar Abdurakhmanov ◽  
Nils Landegren ◽  
Liza Löf ◽  
...  
Keyword(s):  
Drug Target ◽  
Kinase Inhibitors ◽  
Expression Profiles ◽  
Clinical Samples ◽  
Target Engagement ◽  
Rolling Circle Replication ◽  
Target Proteins ◽  
Dna Conjugates

Abstract Drugs are designed to bind their target proteins in physiologically relevant tissues and organs to modulate biological functions and elicit desirable clinical outcomes. Information about target engagement at cellular and subcellular resolution is therefore critical for guiding compound optimization in drug discovery, and for probing resistance mechanisms to targeted therapies in clinical samples. We describe a target engagement-mediated amplification (TEMA) technology, where oligonucleotide-conjugated drugs are used to visualize and measure target engagement in situ, amplified via rolling-circle replication of circularized oligonucleotide probes. We illustrate the TEMA technique using dasatinib and gefitinib, two kinase inhibitors with distinct selectivity profiles. In vitro binding by dasatinib probe to arrays of displayed proteins accurately reproduced known selectivity profiles, while their differential binding to a panel of fixed adherent cells agreed with expectations from expression profiles of the cells. These findings were corroborated by competition experiments using kinase inhibitors with overlapping and non-overlapping target specificities, and translated to pathology tissue sections. We also introduce a proximity ligation variant of TEMA in which these drug-DNA conjugates are combined with antibody-DNA conjugates to selectively investigate binding to specific target proteins of interest. This form of the assay serves to improve resolution of binding to on- and off-target proteins. In conclusion, TEMA has the potential to aid in drug development and clinical routine by conferring valuable insights in drug-target interactions at spatial resolution in protein arrays, cells and tissues.

Structural motifs in drug target proteins of global infectious agents

1999 ◽  
Vol 124 ◽  
pp. S57
Author(s):  
Wim G.J. Hol ◽  
Stephen Suresh ◽  
Jungwoo Choe ◽  
David Chudzik ◽  
Christophe L.M.J. Verlinde
Keyword(s):  
Drug Target ◽  
Infectious Agents ◽  
Structural Motifs ◽  
Target Proteins

scholarly journals Prioritizing and modelling of putative drug target proteins of Candida albicans by system biology approach

2018 ◽  
Vol 65 (2) ◽  
pp. 209-218 ◽  
Author(s):  
Tariq Ismail ◽  
Nighat Fatima ◽  
Syed Aun Muhammad ◽  
Syed Saoud Zaidi ◽  
Nisar Rehman ◽  
...  
Keyword(s):  
Candida Albicans ◽  
Drug Target ◽  
Drug Targets ◽  
Cellular Localization ◽  
Critical Role ◽  
Human Microbiome ◽  
Genomic Analysis ◽  
Cellular Systems ◽  
Gut Flora ◽  
Target Proteins

Candida albicans (C. albicans) is one of the major source of nosocomial infections in human which may prove fatal in 30% of cases. The hospital acquired infection is very difficult to affectively treat due to the presence of drug resistant pathogenic strains, therefore there is a need to find alternative drug targets to cure this infection. In silico and computational level frame work was used to prioritize and establish antifungal drug targets of Candida albicans. The identification of putative drug targets was based on acquiring completely 5090 annotated genes of Candida albicans from available databases which was categorized into essential and non-essential genes. The result indicated 9% proteins were essential that could become potential candidates for intervention which might result in pathogen death. We studied cluster of orthologs and the subtractive genomic analysis of these essential proteins against human genome as a reference to minimize the side effects. It was seen that 14% of Candidal proteins were evolutionary related to the human proteins while 86% are non-human homologs. In next step for the selection of compatible drug targets, the non-human homologs were sequentially compared to human microbiome data to minimize the potential effects against gut flora which accumulated to 38% of essential genome. The sub-cellular localization of these candidate proteins in fungal cellular systems exhibited that 80% are cytoplasmic, 10% are mitochondrial and remaining 10 % are associated with cell wall. The role of these non-human and non-gut flora putative target proteins in Candidal biological pathways was studied and on the basis of their integrated and critical role 4-proteins were selected for molecular modeling.  For drug designing and development, five quality and reliable protein models with more than 70% homology were constructed. Our study will be an effective framework for drug target identifications of pathogenic microbial strains and development of new therapies against these infections.

ChemInform Abstract: Collaborative Use of Neutron and X-Ray for Determination of Drug Target Proteins

ChemInform ◽  
2010 ◽  
Vol 41 (41) ◽  
pp. no-no
Author(s):  
Ryota Kuroki ◽  
Taro Tamada ◽  
Kazuo Kurihara ◽  
Takashi Ohhara ◽  
Motoyasu Adachi
Keyword(s):  
Drug Target ◽  
X Ray ◽  
Target Proteins

scholarly journals An AlphaScreenTM-Based High-Throughput Screen to Identify Inhibitors of Hsp90-Cochaperone Interaction

2009 ◽  
Vol 14 (3) ◽  
pp. 273-281 ◽  
Author(s):  
Fang Yi ◽  
Pingjun Zhu ◽  
Noel Southall ◽  
James Inglese ◽  
Christopher P. Austin ◽  
...  
Keyword(s):  
High Throughput ◽  
Drug Target ◽  
Essential Role ◽  
High Throughput Screen ◽  
Chemical Probes ◽  
Chemical Genomics ◽  
Hsp90 Inhibitors ◽  
Hsp90 Function ◽  
Good Agreement

Hsp90 has emerged as an important anticancer drug target because of its essential role in promoting the folding and maturation of many oncogenic proteins. The authors describe the development of the first high-throughput screen, based on AlphaScreen™ technology, to identify a novel type of Hsp90 inhibitors that interrupt its interaction with the cochaperone HOP. The assay used the 20-mer C-terminal peptide of Hsp90 and the TPR2A domain of HOP. Assay specificity was demonstrated by measuring different interactions using synthetic peptides, with measured IC50s in good agreement with reported values. The assay was stable over 12 h and tolerated DMSO up to 5%. The authors first validated the assay by screening against 20,000 compounds in a 384-well format. After further optimization into a 1536-well format, it was screened against an NIH Chemical Genomics Center library of 76,134 compounds, with a signal-to-background ratio of 78 and Z′ factor of 0.77. The present assay can be used for discovery of novel small-molecule Hsp90 inhibitors that can be used as chemical probes to investigate the role of cochaperones in Hsp90 function. Such molecules have the potential to be developed into novel anticancer drugs, for use alone or in combination with other Hsp90 inhibitors. ( Journal of Biomolecular Screening 2009:273-281)

Towards investigation of the inhibitor-recognition mechanisms of drug-target proteins by neutron crystallography

2010 ◽  
Vol 66 (11) ◽  
pp. 1126-1130 ◽  
Author(s):  
Ryota Kuroki ◽  
Nobuo Okazaki ◽  
Motoyasu Adachi ◽  
Takashi Ohhara ◽  
Kazuo Kurihara ◽  
...  
Keyword(s):  
Drug Target ◽  
Target Proteins ◽  
Inhibitor Recognition ◽  
Neutron Crystallography
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