Re: [ccp4bb] Assays for protein-ligand interaction?
You can try MST(MicroScale Thermophoresis), if there is an instrument available. It is like a mini ITC, but much more versatile. The sample consumption is very low, claimed by the company that sells the instrument. http://www.nanotemper-technologies.com/technology/microscale-thermophoresis/article/key-features/ It is possible to ask the company to perform a demo and test some samples in your lab. Good luck, Jia-Ying On 13 January 2014 14:47, George Kontopidis wrote: > *Specifically for fluorescence* > > does your ligand fluoresce? > > It is possible if it has indol group or some aromatic organic compound > > > > Does your protein has a tryptophan or tyrosines in the binding site? > > If yes may be a fluorescence titration experiment could be the solution. > > > > Also fluorescence needs very low concentration of protein (nM to microM) > > > > george > > > > > > *From:* CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] *On Behalf Of > *Acoot > Brett > *Sent:* Monday, January 13, 2014 3:09 PM > *To:* CCP4BB@JISCMAIL.AC.UK > *Subject:* Re: [ccp4bb] Assays for protein-ligand interaction? > > > > FRET， CD， Fluorescence， NMR chemical shift assay, isotope-labelled ligand > interaction assay, protein melting temperature assay, gel filtration > retention assay, gyration radius assay by Malls, native page gel analysis, > etc. > > > > Acoot > > > > On Monday, 13 January 2014 8:51 PM, HJ Lee wrote: > > Sorry for the off topic. I'm looking for a way to monitor > protein-(potential) ligand interaction. The ligand is small molecule > (mw~250) and we're looking for its potential interaction with couple human > proteins. (We do not know this small molecule interacts with these human > protein or not.) > > > > Is there any efficient way to quickly identify whether this ligand > interacts with those human protein? We can buy some protein, but the amount > of commercially available purified proteins is very little, making them > hard to be analyzed by some good methods (e.g. ITC). > > > > That would be really great if anyone suggest any idea. Sorry for the off > topic question again. > > > > Thanks! > > >
Re: [ccp4bb] Assays for protein-ligand interaction?
Specifically for fluorescence does your ligand fluoresce? It is possible if it has indol group or some aromatic organic compound Does your protein has a tryptophan or tyrosines in the binding site? If yes may be a fluorescence titration experiment could be the solution. Also fluorescence needs very low concentration of protein (nM to microM) george From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of Acoot Brett Sent: Monday, January 13, 2014 3:09 PM To: CCP4BB@JISCMAIL.AC.UK Subject: Re: [ccp4bb] Assays for protein-ligand interaction? FRET， CD， Fluorescence， NMR chemical shift assay, isotope-labelled ligand interaction assay, protein melting temperature assay, gel filtration retention assay, gyration radius assay by Malls, native page gel analysis, etc. Acoot On Monday, 13 January 2014 8:51 PM, HJ Lee < <mailto:hojunle...@gmail.com> hojunle...@gmail.com> wrote: Sorry for the off topic. I'm looking for a way to monitor protein-(potential) ligand interaction. The ligand is small molecule (mw~250) and we're looking for its potential interaction with couple human proteins. (We do not know this small molecule interacts with these human protein or not.) Is there any efficient way to quickly identify whether this ligand interacts with those human protein? We can buy some protein, but the amount of commercially available purified proteins is very little, making them hard to be analyzed by some good methods (e.g. ITC). That would be really great if anyone suggest any idea. Sorry for the off topic question again. Thanks!
Re: [ccp4bb] Assays for protein-ligand interaction?
FRET， CD， Fluorescence， NMR chemical shift assay, isotope-labelled ligand interaction assay, protein melting temperature assay, gel filtration retention assay, gyration radius assay by Malls, native page gel analysis, etc. Acoot On Monday, 13 January 2014 8:51 PM, HJ Lee wrote: Sorry for the off topic. I'm looking for a way to monitor protein-(potential) ligand interaction. The ligand is small molecule (mw~250) and we're looking for its potential interaction with couple human proteins. (We do not know this small molecule interacts with these human protein or not.) Is there any efficient way to quickly identify whether this ligand interacts with those human protein? We can buy some protein, but the amount of commercially available purified proteins is very little, making them hard to be analyzed by some good methods (e.g. ITC). That would be really great if anyone suggest any idea. Sorry for the off topic question again. Thanks!
Re: [ccp4bb] Assays for protein-ligand interaction?
Ho Jun Lee, Have you thought about differential scanning fluorimetry (Thermofluor)? With this biophysical technique you can characterize protein-ligand interactions and screen a wide variety of ligands using minimal concentration of ligand and protein. All you need is a quantitative PCR machine (qPCR). Here is a reference for you https://chemistry.osu.edu/~magliery/pdfs/LavinderMagliery2009JACS.pdf Lorenzo Ihsan Finci, Ph.D.Postdoctoral Scientist, Wang LaboratoryHarvard Medical SchoolDana-Farber Cancer InstituteBoston, MA Peking UniversityCollege of Life SciencesBeijing, China Date: Mon, 13 Jan 2014 21:50:07 +0900 From: hojunle...@gmail.com Subject: [ccp4bb] Assays for protein-ligand interaction? To: CCP4BB@JISCMAIL.AC.UK Sorry for the off topic. I'm looking for a way to monitor protein-(potential) ligand interaction. The ligand is small molecule (mw~250) and we're looking for its potential interaction with couple human proteins. (We do not know this small molecule interacts with these human protein or not.) Is there any efficient way to quickly identify whether this ligand interacts with those human protein? We can buy some protein, but the amount of commercially available purified proteins is very little, making them hard to be analyzed by some good methods (e.g. ITC). That would be really great if anyone suggest any idea. Sorry for the off topic question again. Thanks!
