Re: [ccp4bb] Strategies for increasing Trp content of proteins

[Artem Evdokimov]

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.  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: 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
>



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/

[ccp4bb] Strategies for increasing Trp content of proteins

[Tanner, John J.]

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: 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

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/

[ccp4bb] 4-year PhD position in Bergen, Norway

[Petri Kursula]

Hi,

we have an opening for a fully funded 4-year PhD position in my group at the 
Unviersity of Bergen, Norway. The position is funded by a recent grant from the 
Norwegian Research Council, and the broad topic is the assembly and properties 
of multilayered proteolipid membranes. 

Please see the full ad below for details. Informal queries are of course 
welcome, but applications must be made through the link in the advert. Note 
that all required documents must be submitted for the application to be 
evaluated.

https://www.jobbnorge.no/en/available-jobs/job/221160/phd-position-4-years 
 

Best regards,
Petri

Petri Kursula
--
Professor 
--
Department of Biomedicine
University of Bergen, Norway
https://link.uib.no/petri 
petri.kurs...@uib.no 
--
Faculty of Biochemistry and Molecular Medicine
Biocenter Oulu
University of Oulu, Finland
--


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/

[ccp4bb] PDRA in software development at York...

[Kevin Cowtan]

Hi all

We have a position available at York to develop web based tools for
molecular graphics and model building. For details, see this page

.

-- 
Professor Kevin Cowtan
Email:  kevin.cow...@york.ac.uk
Pronouns:   Please use they/them when referring to me in professional
contexts
ORCiD:  -0002-0189-1437 
Disclaimer: http://w 
ww.york.ac.uk/docs/disclaimer/email.htm




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/

[ccp4bb] CCP-EM Spring Symposium VIII 2022 3-5th May 2022

[Tom Burnley - STFC UKRI]

Dear CCP-EM,

CCP-EM are pleased to announce the eighth annual 'CCP-EM Spring Symposium', 
3-5th May 2022.  The conference aims to provide a forum to highlight state of 
the art developments in computational cryoEM and related themes as well as 
showcasing outstanding recent biological applications.  We aim to promote an 
inclusive, friendly atmosphere welcoming both old and new to the community.  
Topics include instrument technology, sample preparation, image processing, 
single particle reconstruction, tomography and model building.  The schedule 
will also include the Diamond Light Source biological cryo-imaging user meeting 
for eBIC & B24.

The conference will be held as a hybrid meeting with the physical event at the 
EMCC in Nottingham and online via Zoom and is kindly sponsored by SPT Labtech 
and Quantifoil.

Registration is open now:
https://cvent.me/449Ozw

More details and schedule are available here:
https://www.ccpem.ac.uk/training/spring_symposium_2022/spring_symposium.php

Confirmed speakers include:
Becky Thompson (Leeds)
Christiane Berger-Schaffitzel (Bristol)
Dari Kimanius (MRC-LMB)
Dorothee Liebschner (Berkeley Lab)
Florian Schur (IST-Austria)
Giulia Zanetti (Birkbeck)
Goedele Maertens (Imperial)
Holger Stark (MPI Biophysical Chemistry)
Katerina Naydenova (MRC-LMB)
Peter Rosenthal (Crick)
Ruben Sanchez-Garcia (Oxford)
Tim Grant (UW-Madison)
Tom Deegan (Edinburgh)
Ulrich Lorenz (EPFL)
Wim Hagen (EMBL)
What’s new at CCP-EM, EBI, RFI and eBIC

Scientific organisers:
Doryen Bubeck (Imperial)
Paula da Fonseca (Glasgow)

Best wishes from the CCP-EM team

This email and any attachments are intended solely for the use of the named 
recipients. If you are not the intended recipient you must not use, disclose, 
copy or distribute this email or any of its attachments and should notify the 
sender immediately and delete this email from your system. UK Research and 
Innovation (UKRI) has taken every reasonable precaution to minimise risk of 
this email or any attachments containing viruses or malware but the recipient 
should carry out its own virus and malware checks before opening the 
attachments. UKRI does not accept any liability for any losses or damages which 
the recipient may sustain due to presence of any viruses.




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/

[ccp4bb] Boehringer Ingelheim is looking for new talents!

[Nico Komarowsky]

Boehringer Ingelheim is currently looking for a "Post Doc Integral Membrane 
Protein (Structural Biology)" (Ref. 220769) located in Biberach (Riß), our 
biggest R&D site in Germany.
You can find the most important information below:
THE POSITION
Take your chance to work with a state-of-the-art-structural biology group with 
dedicated protein biochemistry-, biophysics-, NMR-, protein crystallography- 
and cryo-EM capabilities.

You are a Structural Biologist with in-depth knowledge of molecular biology, 
protein biochemistry and biophysics as well as a strong interest to work in 
drug discovery? If you want to work for a company that changes lives for the 
better, join our team within our Medicinal Chemistry department at our biggest 
research site!

Discover our Biberach site: xplorebiberach.com

Tasks & responsibilities

  *   You develop and evaluate approaches for the structure determination of 
medically highly relevant human membrane proteins.
  *   Designing expression constructs and screening expression and purification 
conditions is also part of your profile.
  *   You use biophysical methods for membrane protein characterization, 
including small-molecule binding.
  *   You determine 3D structures using state-of-the-art techniques (e.g., 
X-ray or cryo-EM)
  *   Take the opportunity to collaborate with experts in the field (internal / 
external), present your results at internal and external meetings and publish 
in peer-reviewed journals.

Requirements

  *   PhD in Structural Biology, Biochemistry, or a related field
  *   Expert knowledge in Structural Biology; cryo-EM expertise is a plus
  *   Expertise in molecular biology as well as in working with membrane 
protein expression in insect- or mammalian cell systems
  *   Skills in the purification and characterization of integral membrane 
proteins
  *   Excellent interpersonal skills paired with the ability to effectively 
interact with stakeholders, both internally and externally
  *   Fluency in English (written and spoken)

You can find further information and apply by following this link: 
https://tas-boehringer.taleo.net/careersection/jobdetail.ftl?job=220769&lang=de



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/

[ccp4bb] PDBe updated mmCIF files now available via FTP

[David Armstrong]

Dear Colleagues,

PDBe has madePDBx/mmCIF files, containing SIFTS mapping information and 
additional, standardised metadata available directly from the EMBL-EBI 
FTP area, from:

https://ftp.ebi.ac.uk/pub/databases/msd/updated_mmcif/

PDBe’s release process creates these enriched PDBx/mmCIF files from PDB 
archive files, containing more consistent, standardised metadata, 
without altering core PDB information, such as atomic coordinates and 
experimental data. These PDBx/mmCIF files have recently been further 
enriched with data from the SIFTS process 
, 
providing improved mapping between structure and sequence data.


These enriched PDBx/mmCIF files are compressed and distributed from two 
subfolders in the FTP area. Firstly, you can efficiently access files 
for individual entries via the ‘divided’ folder, where each entry is 
placed within a folder identified by the second and third characters of 
the PDB entry ID (e.g. divided/at/1atp_updated.cif.gz). Alternatively, 
all the compressed updated PDBx mmCIF files in the PDB archive are 
available from the ‘all’ folder (e.g. all/1atp_updated.cif.gz).


Alternatively, these files can also be downloaded from PDBe entry pages 
(https://www.ebi.ac.uk/pdbe/entry-files/download/7o9f_updated.cif). 
These files are also accessible via static URL 
(https://www.ebi.ac.uk/pdbe/static/entry/7o9f_updated.cif) or via PDBe 
bulk download service available at https://www.ebi.ac.uk/pdbe/download/api.


If you have any queries about these files or how to access them, please 
contact our helpdesk at pdb...@ebi.ac.uk.


Kind Regards,
David Armstrong

--
David Armstrong
Outreach and Training Coordinator
PDBe
European Bioinformatics Institute (EMBL-EBI)
European Molecular Biology Laboratory
Wellcome Trust Genome Campus
Hinxton
Cambridge CB10 1SD UK
Tel: +44 1223 492544



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/

[ccp4bb] post-doc and PhD position at KU Leuven, ligand-gated ion channels

[Chris Ulens]

This is a kind reminder that the positions of post-doc and PhD in our lab are 
still open for applicants.

Best regards
Chris


> On 18 Jan 2022, at 16:20, Chris Ulens  wrote:
> 
> Dear members of the community,
> One post-doctoral position and one PhD position are available in the 
> Laboratory of Prof. Chris Ulens, KU Leuven, Belgium (http://www.xtal.be 
> ). The research in the Laboratory of Structural 
> Neurobiology is aimed at understanding the molecular mechanism of 
> ligand-gated ion channels. Our methods rely on X-ray crystallography and 
> cryo-EM in combination with electrophysiological recordings and 
> fluorescence-based techniques. Our laboratory hosts an integrated 
> macromolecular crystallization facility with a Mosquito crystallization robot 
> and two RockImagers. We have access to a 200 kV Glacios microscope on campus 
> as well as 300 kV JEOL CryoARM300 at the nearby EM facility in Brussels. 
> Current projects are situated at the crossroads of structural elucidation and 
> drug discovery of different pentameric ligand-gated ion channels.
> 
> We would like to recruit a post-doc and PhD student with prior experience in 
> purification, cryo-EM or X-ray crystallography of membrane proteins. The 
> position is open immediately and applications will be accepted until the 
> position is filled. Applicants are requested to submit a CV with publication 
> list and two references. E-mail to chris.ul...@kuleuven.be 
> 
> 
> 
> Selection of recent publications:
> 
> 
> A lipid site shapes the agonist response of a pentameric ligand-gated ion 
> channel. Hénault CM, Govaerts C, Spurny R, Brams M, Estrada-Mondragon A, 
> Lynch J, Bertrand D, Pardon E, Evans GL, Woods K, Elberson BW, Cuello LG, 
> Brannigan G, Nury H, Steyaert J, Baenziger JE, Ulens C. Nat Chem Biol. 
> 2019;15(12):1156-1164. doi: 10.1038/s41589-019-0369-4.
> 
> 
> Modulation of the Erwinia ligand-gated ion channel (ELIC) and the 5-HT3 
> receptor via a common vestibule site. Brams M, Govaerts C, Kambara K, Price 
> KL, Spurny R, Gharpure A, Pardon E, Evans GL, Bertrand D, Lummis SC, Hibbs 
> RE, Steyaert J, Ulens C. Elife. 2020;9:e51511. doi: 10.7554/eLife.51511.
> 
> 
> The structural basis for an on-off switch controlling Gβγ-mediated inhibition 
> of TRPM3 channels. Behrendt M, Gruss F, Enzeroth R, Dembla S, Zhao S, 
> Crassous PA, Mohr F, Nys M, Louros N, Gallardo R, Zorzini V, Wagner D, 
> Economou A, Rousseau F, Schymkowitz J, Philipp SE, Rohacs T, Ulens C, 
> Oberwinkler J. Proc Natl Acad Sci U S A. 2020;117(46):29090-29100. doi: 
> 10.1073/pnas.2001177117.
> 
> 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/


smime.p7s
Description: S/MIME cryptographic signature