Yes we chose the sites based on predicted or known structure, to be sure. No that was not a published thing, curse of commercial work :(
Happy to help on a private channel. Artem On Sat, Feb 19, 2022, 1:42 PM Tanner, John J. <[email protected]> wrote: > Casper: You make a good point about using 205 nm. I checked our system > this morning and the wavelength is fixed at 280 nm (U9-L). > > > > Artem: Did you consider secondary structure and/or amino acid residue type > when choosing sites for mutagenesis? Is your work published? > > > > > > *From: *CCP4 bulletin board <[email protected]> on behalf of Casper > Wilkens <[email protected]> > *Date: *Saturday, February 19, 2022 at 2:19 AM > *To: *[email protected] <[email protected]> > *Subject: *[ccp4bb] Sv: [ccp4bb] Strategies for increasing Trp content of > proteins > > *WARNING:* This message has originated from an External Source. This may > be a phishing expedition that can result in unauthorized access to our IT > System. Please use proper judgment and caution when opening attachments, > clicking links, or responding to this email. > > Is absorbance at 205 nm not a possibility, Jack? > > > > *Casper Wilkens * > > Asst. Prof. > > Structural Enzymology & Biorefining > > DTU Bioengineering > > > > *Technical University of Denmark* > > Department of Biotechnology and Biomedicine > > Søltofts Plads > Building 224 > Room 028 > > 2800 Kgs. Lyngby > > [email protected] <[email protected]> > > www.bioengineering.dtu.dk/ > <https://nam02.safelinks.protection.outlook.com/?url=http%3A%2F%2Fwww.bioengineering.dtu.dk%2F&data=04%7C01%7Ctannerjj%40MISSOURI.EDU%7Ccd1aefc8cc284ea9e44d08d9f38074db%7Ce3fefdbef7e9401ba51a355e01b05a89%7C0%7C0%7C637808555459123981%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C3000&sdata=viaDJTf6EgVVTGxk4AGAWEyluluwh9rtnwUCUUULmo0%3D&reserved=0> > > https://www.cazypedia.org/index.php/User:Casper_Wilkens > <https://nam02.safelinks.protection.outlook.com/?url=https%3A%2F%2Fwww.cazypedia.org%2Findex.php%2FUser%3ACasper_Wilkens&data=04%7C01%7Ctannerjj%40MISSOURI.EDU%7Ccd1aefc8cc284ea9e44d08d9f38074db%7Ce3fefdbef7e9401ba51a355e01b05a89%7C0%7C0%7C637808555459123981%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C3000&sdata=yJVCEDisnY3LmyvCPPjFKmybTNFrQHlEKjrEmQPVtHE%3D&reserved=0> > https://twitter.com/protein_artist > <https://nam02.safelinks.protection.outlook.com/?url=https%3A%2F%2Ftwitter.com%2Fprotein_artist&data=04%7C01%7Ctannerjj%40MISSOURI.EDU%7Ccd1aefc8cc284ea9e44d08d9f38074db%7Ce3fefdbef7e9401ba51a355e01b05a89%7C0%7C0%7C637808555459123981%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C3000&sdata=bKeX%2FhUL7pnDA3DxL5ZFcTUlHkL5x2Vd7VsrJNPA0H0%3D&reserved=0> > https://www.researchgate.net/profile/Casper_Wilkens > <https://nam02.safelinks.protection.outlook.com/?url=https%3A%2F%2Fwww.researchgate.net%2Fprofile%2FCasper_Wilkens&data=04%7C01%7Ctannerjj%40MISSOURI.EDU%7Ccd1aefc8cc284ea9e44d08d9f38074db%7Ce3fefdbef7e9401ba51a355e01b05a89%7C0%7C0%7C637808555459123981%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C3000&sdata=VuBOXy%2BOSmA6usq%2BahiVlN2%2Fm19%2FnlVEqNXDRMRnrkY%3D&reserved=0> > ------------------------------ > > *Fra:* CCP4 bulletin board <[email protected]> på vegne af Artem > Evdokimov <[email protected]> > *Sendt:* 19. februar 2022 02:12:50 > *Til:* [email protected] > *Emne:* Re: [ccp4bb] Strategies for increasing Trp content of proteins > > > > We have had success placing single trp on the outside of proteins, as > weird as it sounds it works pretty well. If your protein is easy to express > you can try a few choices. > > > > If your protein is difficult to express, a fusion with a trp-positive > domain or with gfp may be a better option. > > > > There are many other options but their application is best considered with > more information in hand > > > > Artem > > > > On Fri, Feb 18, 2022, 8:02 PM Tanner, John J. <[email protected]> > wrote: > > Dear CCP4BB, > > We are working on a protein that has no Trp residues, which makes > chromatography challenging due to the low absorbance at 280 nm (Abs 0.1% = > 0.104). Does anyone have experience using mutagenesis to increase the Trp > content of proteins? > > Thanks, > > Jack Tanner > > > > -- > > John J. Tanner > > Professor of Biochemistry and Chemistry > > Associate Chair of Biochemistry > > Department of Biochemistry > > University of Missouri > 117 Schweitzer Hall > > 503 S College Avenue > Columbia, MO 65211 > Phone: 573-884-1280 > > Email: [email protected] <[email protected]> > https://cafnrfaculty.missouri.edu/tannerlab/ > > Lab: Schlundt Annex rooms 3,6,9, 203B, 203C > > Office: Schlundt Annex 203A > > > > > > > ------------------------------ > > To unsubscribe from the CCP4BB list, click the following link: > https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 > <https://nam02.safelinks.protection.outlook.com/?url=https%3A%2F%2Fwww.jiscmail.ac.uk%2Fcgi-bin%2FWA-JISC.exe%3FSUBED1%3DCCP4BB%26A%3D1&data=04%7C01%7Ctannerjj%40MISSOURI.EDU%7Ccd1aefc8cc284ea9e44d08d9f38074db%7Ce3fefdbef7e9401ba51a355e01b05a89%7C0%7C0%7C637808555459123981%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C3000&sdata=lqUvJHHOWqoIPD4zzPwhjINYcanSB1R4e97nPfeBiSA%3D&reserved=0> > > > ------------------------------ > > To unsubscribe from the CCP4BB list, click the following link: > https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 > <https://nam02.safelinks.protection.outlook.com/?url=https%3A%2F%2Fwww.jiscmail.ac.uk%2Fcgi-bin%2FWA-JISC.exe%3FSUBED1%3DCCP4BB%26A%3D1&data=04%7C01%7Ctannerjj%40MISSOURI.EDU%7Ccd1aefc8cc284ea9e44d08d9f38074db%7Ce3fefdbef7e9401ba51a355e01b05a89%7C0%7C0%7C637808555459123981%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C3000&sdata=lqUvJHHOWqoIPD4zzPwhjINYcanSB1R4e97nPfeBiSA%3D&reserved=0> > > > ------------------------------ > > To unsubscribe from the CCP4BB list, click the following link: > https://www.jiscmail.ac.uk/cgi-bin/WA-JISC.exe?SUBED1=CCP4BB&A=1 > <https://nam02.safelinks.protection.outlook.com/?url=https%3A%2F%2Fwww.jiscmail.ac.uk%2Fcgi-bin%2FWA-JISC.exe%3FSUBED1%3DCCP4BB%26A%3D1&data=04%7C01%7Ctannerjj%40MISSOURI.EDU%7Ccd1aefc8cc284ea9e44d08d9f38074db%7Ce3fefdbef7e9401ba51a355e01b05a89%7C0%7C0%7C637808555459123981%7CUnknown%7CTWFpbGZsb3d8eyJWIjoiMC4wLjAwMDAiLCJQIjoiV2luMzIiLCJBTiI6Ik1haWwiLCJXVCI6Mn0%3D%7C3000&sdata=lqUvJHHOWqoIPD4zzPwhjINYcanSB1R4e97nPfeBiSA%3D&reserved=0> > > ------------------------------ > > 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/
