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 <[email protected]> on behalf of Gloria Borgstahl <[email protected]> Sent: Tuesday, September 22, 2020 2:28 PM To: [email protected] <[email protected]> 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 <[email protected]<mailto:[email protected]>> 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 [email protected]<mailto:[email protected]> (214) 645-6383 (phone) (214) 645-6353 (fax) On Sep 22, 2020, at 12:08 PM, Srivastava, Dhiraj <[email protected]<mailto:[email protected]>> 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. Thank you Dhiraj ________________________________ To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 CAUTION: This email originated from outside UTSW. Please be cautious of links or attachments, and validate the sender's email address before replying. ________________________________ UT Southwestern Medical Center The future of medicine, today. ________________________________ To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 ________________________________ To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 This message was issued to members of www.jiscmail.ac.uk/CCP4BB, a mailing list hosted by www.jiscmail.ac.uk, terms & conditions are available at https://www.jiscmail.ac.uk/policyandsecurity/
