scholarly journals Multiplexed Screening of Thousands of Natural Products for Protein–Ligand Binding in Native Mass Spectrometry

2021 ◽  
Author(s):  
Giang T. H. Nguyen ◽  
Jack L. Bennett ◽  
Sherrie Liu ◽  
Sarah E. Hancock ◽  
Daniel L. Winter ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Natural Products ◽  
Ligand Binding ◽  
Native Mass Spectrometry

scholarly journals Unpicking allosteric mechanisms of homo-oligomeric proteins by determining their successive ligand binding constants

2018 ◽  
Vol 373 (1749) ◽  
pp. 20170176 ◽  
Author(s):  
Ranit Gruber ◽  
Amnon Horovitz
Keyword(s):  
Mass Spectrometry ◽  
Ligand Binding ◽  
Single Molecule ◽  
Binding Constants ◽  
Molecular Machines ◽  
Native Mass Spectrometry ◽  
Oligomeric Proteins ◽  
Allosteric Mechanisms ◽  
The Relationship

Advances in native mass spectrometry and single-molecule techniques have made it possible in recent years to determine the values of successive ligand binding constants for large multi-subunit proteins. Given these values, it is possible to distinguish between different allosteric mechanisms and, thus, obtain insights into how various bio-molecular machines work. Here, we describe for ring-shaped homo-oligomers, in particular, how the relationship between the values of successive ligand binding constants is diagnostic for concerted, sequential and probabilistic allosteric mechanisms. This article is part of a discussion meeting issue ‘Allostery and molecular machines’.

scholarly journals Multiplexed screening of thousands of natural products for protein-ligand binding in native mass spectrometry

2021 ◽  
Author(s):  
Giang Nguyen ◽  
Jack Bennett ◽  
Sherrie Liu ◽  
Sarah Hancock ◽  
Daniel Winter ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Natural Products ◽  
Drug Discovery ◽  
Small Molecules ◽  
Natural Product ◽  
Drug Targets ◽  
Structural Diversity ◽  
Native Mass Spectrometry ◽  
Protein Targets ◽  
Rapid Measurement

The structural diversity of natural products offers unique opportunities for drug discovery, but challenges associated with their isolation and screening can hinder the identification of drug-like molecules from complex natural product extracts. Here we introduce a mass spectrometry-based approach that integrates untargeted metabolomics with multistage, high-resolution native mass spectrometry to rapidly identify natural products that bind to therapeutically relevant protein targets. By directly screening crude natural product extracts containing thousands of drug-like small molecules using a single, rapid measurement, novel natural product ligands of human drug targets could be identified without fractionation. This method should significantly increase the efficiency of target-based natural product drug discovery workflows.

Multiplexed screening of thousands of natural products for protein-ligand binding in native mass spectrometry

2021 ◽  
Author(s):  
Giang Nguyen ◽  
Jack Bennett ◽  
Sherrie Liu ◽  
Sarah Hancock ◽  
Daniel Winter ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Natural Products ◽  
Drug Discovery ◽  
Small Molecules ◽  
Natural Product ◽  
Drug Targets ◽  
Structural Diversity ◽  
Native Mass Spectrometry ◽  
Protein Targets ◽  
Rapid Measurement

The structural diversity of natural products offers unique opportunities for drug discovery, but challenges associated with their isolation and screening can hinder the identification of drug-like molecules from complex natural product extracts. Here we introduce a mass spectrometry-based approach that integrates untargeted metabolomics with multistage, high-resolution native mass spectrometry to rapidly identify natural products that bind to therapeutically relevant protein targets. By directly screening crude natural product extracts containing thousands of drug-like small molecules using a single, rapid measurement, novel natural product ligands of human drug targets could be identified without fractionation. This method should significantly increase the efficiency of target-based natural product drug discovery workflows.

scholarly journals Determination of Antiplasmodial Activity and Binding Affinity of Selected Natural Products towards PfTrxR and PfGR

2013 ◽  
Vol 8 (8) ◽  
pp. 1934578X1300800
Author(s):  
Ranjith Munigunti ◽  
Katja Becker ◽  
Reto Brun ◽  
Angela I. Calderón
Keyword(s):  
Mass Spectrometry ◽  
Natural Products ◽  
Ligand Binding ◽  
Binding Assay ◽  
Antimalarial Activity ◽  
Antiplasmodial Activity ◽  
Binding Affinities ◽  
Ligand Binding Assay

In our study, the binding affinities of selected natural products towards PfTrxR, PfGR, human TrxR and human GR were determined using a mass spectrometry based ligand binding assay. The in vitro antimalarial activity and cytotoxicity of these ligands were also determined. Catharanthine, 11-(OH)-coronaridine, hernagine, vobasine and hispolone displayed antiplasmodial activity against PfK1 (IC50 = 0.996–3.63 μg/mL).

Mass spectrometry tools for screening of marine cyanobacterial natural products

Planta Medica ◽  
2015 ◽  
Vol 81 (11) ◽  
Author(s):  
T Luzzatto Knaan ◽  
N Garg ◽  
Y Peng ◽  
T Alexandrov ◽  
G Navarro ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Natural Products

Applications of ambient mass spectrometry of natural products in the fields of food safety, phytochemistry and forensics

Planta Medica ◽  
2016 ◽  
Vol 81 (S 01) ◽  
pp. S1-S381
Author(s):  
TA van Beek ◽  
W Duvivier ◽  
Y Shen ◽  
B Chen ◽  
MWF Nielen
Keyword(s):  
Mass Spectrometry ◽  
Natural Products ◽  
Food Safety ◽  
Ambient Mass Spectrometry

Rapid Online Buffer Exchange: A Method for Screening of Proteins, Protein Complexes, and Cell Lysates by Native Mass Spectrometry

2019 ◽  
Author(s):  
Zachary VanAernum ◽  
Florian Busch ◽  
Benjamin J. Jones ◽  
Mengxuan Jia ◽  
Zibo Chen ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
High Speed ◽  
Structural Information ◽  
Protein Complexes ◽  
High Sensitivity ◽  
Native Mass Spectrometry ◽  
Structural Features ◽  
Consumer Products ◽  
Cell Lysates ◽  
Protein Expression And Purification

It is important to assess the identity and purity of proteins and protein complexes during and after protein purification to ensure that samples are of sufficient quality for further biochemical and structural characterization, as well as for use in consumer products, chemical processes, and therapeutics. Native mass spectrometry (nMS) has become an important tool in protein analysis due to its ability to retain non-covalent interactions during measurements, making it possible to obtain protein structural information with high sensitivity and at high speed. Interferences from the presence of non-volatiles are typically alleviated by offline buffer exchange, which is timeconsuming and difficult to automate. We provide a protocol for rapid online buffer exchange (OBE) nMS to directly screen structural features of pre-purified proteins, protein complexes, or clarified cell lysates. Information obtained by OBE nMS can be used for fast (<5 min) quality control and can further guide protein expression and purification optimization.

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