Hi Jack, We routinely include a single glycine downstream of the SUMO (so, …QIGGG). This essentially always gives good cleavage with UD, assuming the construct isn’t aggregated or otherwise unhappy.
Re the His tag: Presumably you mean at the N-terminus of SUMO? In this case we’ve had good luck with MGHHHHHHG. If I was going to redesign this from scratch, I might make it a His-10 tag, but that’s really just a theoretical notion; practically speaking, we’ve never had the impetus to change things. Details in PMCID PMC1892228. Cheers, Pat --------------------------------------------------------------------------------------- Patrick J. Loll, Ph. D. (he, him, his) Professor of Biochemistry & Molecular Biology Drexel University College of Medicine Room 10-102 New College Building 245 N. 15th St., Mailstop 497 Philadelphia, PA 19102 USA (215) 762-7706 pjl...@gmail.com pj...@drexel.edu > On 27 Apr 2021, at 4:00 PM, Tanner, John J. <tanne...@missouri.edu> wrote: > > Sorry for the non-CCP4 question… > > I would like ask your advice on designing SUMO fusion proteins. Specifically, > is it recommended to include a linker between the C-terminus of SUMO (-QIGG) > and the N-terminus of the target protein? Figure 3 of Guerrero et al. > (https://pubmed.ncbi.nlm.nih.gov/26297996/) suggests that including a linker > of SGG or SGGG greatly improves cleavage efficiency by Ulp1. However, the > often-stated advantage of SUMO is that, in the absence of a linker, the > target protein contains no extra residues after cleavage. > > I’ll also take any advice on your favorite N-terminal His tag for SUMO > fusions. I am considering MGSSHHHHHHG, based on two Acta F papers > (https://pubmed.ncbi.nlm.nih.gov/22949189/, > https://pubmed.ncbi.nlm.nih.gov/18391421/), or perhaps MGSSHHHHHHSSG, which > is from pET14b. > > Thanks, > > Jack > > > -- > 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: tanne...@missouri.edu > 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 > Patrick Loll pjl...@gmail.com ######################################################################## 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/
