Hi Dhiraj, you could also consider making a fusion protein of your protein with itself, with a suitable long linker (gly-ser-gly-ser etc.?) in between. At least that dimer won't dissociate. Best, Herman
Von: CCP4 bulletin board <CCP4BB@JISCMAIL.AC.UK> Im Auftrag von Srivastava, Dhiraj Gesendet: Mittwoch, 23. September 2020 21:39 An: CCP4BB@JISCMAIL.AC.UK Betreff: [EXTERNAL] Re: [ccp4bb] dimeric tag to induce the homodimerization of protein EXTERNAL : Real sender is owner-ccp...@jiscmail.ac.uk<mailto:owner-ccp...@jiscmail.ac.uk> Thanks everyone for all the nice suggestions. regarding Matthew's question, Yes, There is a possibility of polydispersity due to the proteins making chain/aggregate, however the affinity between the dimeric protein A and monomeric protein B is poor and kinetics of dissociation is very fast thus complex can not survive gel filtration chromatography. So, by increasing the avidity, We are hoping that only 2:2 complex will survive the gel filtration and we will be able to isolate monodispersed complex for further biophysical studies like SAXS or crystallization. Thank you Dhiraj ________________________________ From: CCP4 bulletin board <CCP4BB@JISCMAIL.AC.UK<mailto:CCP4BB@JISCMAIL.AC.UK>> on behalf of Gloria Borgstahl <gborgst...@gmail.com<mailto:gborgst...@gmail.com>> Sent: Tuesday, September 22, 2020 2:28 PM To: CCP4BB@JISCMAIL.AC.UK<mailto:CCP4BB@JISCMAIL.AC.UK> <CCP4BB@JISCMAIL.AC.UK<mailto:CCP4BB@JISCMAIL.AC.UK>> Subject: Re: [ccp4bb] dimeric tag to induce the homodimerization of protein I was thinking the form of GFP that dimerizes. This would also make it easy to track where the protein is. On Tue, Sep 22, 2020 at 1:28 PM Diana Tomchick <diana.tomch...@utsouthwestern.edu<mailto:diana.tomch...@utsouthwestern.edu>> wrote: Any dimeric tag should work if you add a long enough linker to satisfy your distance criterion. GST, for example. Download the coordinates and get a rough idea how long the linker would have to be for your protein. Diana ************************************************** Diana R. Tomchick Professor Departments of Biophysics and Biochemistry UT Southwestern Medical Center 5323 Harry Hines Blvd. Rm. ND10.214A Dallas, TX 75390-8816 diana.tomch...@utsouthwestern.edu<mailto:diana.tomch...@utsouthwestern.edu> (214) 645-6383 (phone) (214) 645-6353 (fax) On Sep 22, 2020, at 12:08 PM, Srivastava, Dhiraj <dhiraj-srivast...@uiowa.edu<mailto:dhiraj-srivast...@uiowa.edu>> wrote: EXTERNAL MAIL Hi I want to make my protein dimeric to increase its affinity for its interaction partner which is a dimer. does anyone know a suitable tag/fusion protein which can be used as C terminal fusion for this purpose? I can not use any of the leucine zipper as I am looking for the distance between the c terminus to be around 30-40 A. 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