20.109(S20):Module 1
Contents
Module 1
Lecturer: Angela Koehler
Instructors: Noreen Lyell, Leslie McClain
, Becky Meyer
TAs:
Lab manager: Hsinhwa Lee
Overview
Chemical probes, or ligands, are important research tools used to explore cellular processes and therapeutic targets. The use of high-throughput and unbiased strategies to identify small molecules that bind specific biomolecules, such as proteins, can provide insight on the structure or function of targets. Additionally, a small-molecule screen can identify new chemical probes for target proteins of interest.
The small-molecule microarray (SMM) is a high-throughput method that enables the detection of protein-ligand binding. Briefly, ligands are 'printed' onto a slide and incubated with purifed protein. Unbound protein is washed from the slide and bound protein is detected using a tag on the protein of interest. Because the location of every ligand on the slide is known, the detection of protein indicates that it is bound to the ligand at that location.
In your experiment, you will use SMM data collected by students from the Sp17 semester to identify ligands that bind to TDP43, a ...
This module has been developed thanks to the generous time and thoughtful efforts of several Koehler Laboratory members, in particular Rob Wilson.
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Lab links: day by day
M1D1: Complete in silico cloning and induce TDP43 protein expression
M1D2: Purify TDP43 protein
M1D3: Assess purity and concentration of TDP43 protein
M1D4: Perform small molecule microarray (SMM) with TDP43 protein
M1D5: Scan SMM slides to identify binders of TDP43 protein
M1D6: Analyze SMM data
M1D7: Examine TDP43 binders for chemical features
Assignments
Data summary
Mini-presentation
References
- A method for the covalent capture and screening of diverse small molecules in a microarray format. Nature Protocols. 1:2344-2352.
- Recent discoveries and applications involving small-molecule microarrays. Chemical Biology. 18:21-28.
