[ccp4bb] AW: [ccp4bb] His-6 versus His-10 tag

[Herman . Schreuder]

Dear everybody who reacted,

It seems that when the His6 construct crystallizes, there is a fair chance that 
the his10 construct will crystallize as well. However, as usual in 
crystallography there is no guarantee, so I have added a TEV site to the 
construct in order to be able to remove the tag if necessary.

Thank you all very much!
Herman

Von: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] Im Auftrag von Savvas 
Savvides
Gesendet: Dienstag, 19. September 2017 16:12
An: CCP4BB@JISCMAIL.AC.UK
Betreff: Re: [ccp4bb] His-6 versus His-10 tag

Hi Herman
the survey article by Carson in Acta D
DOI:

10.1107/S0907444906052024<https://urldefense.proofpoint.com/v2/url?u=https-3A__doi.org_10.1107_S0907444906052024=DwMFAg=Dbf9zoswcQ-CRvvI7VX5j3HvibIuT3ZiarcKl5qtMPo=HK-CY_tL8CLLA93vdywyu3qI70R4H8oHzZyRHMQu1AQ=f0yyUbNahC99mMQ7mNgibBIjHQnTAYPdgDT1unpORi8=0ue_qfHHBCOr_J006W2KUHZYoPsjHWboidAAqLjps8g=>
from a decade ago may provide relevant information/insights.
best wishes
Savvas




On 19 Sep 2017, at 12:10, 
herman.schreu...@sanofi.com<mailto:herman.schreu...@sanofi.com> wrote:

Dear BB,

We are planning the production of a protein for crystallization. From 
literature, we know that the construct with a 6-histidine tag crystallizes. 
However, for other biophysical measurements, we would prefer to have a 
10-histidine tag.

Does anyone has experience with His-6 versus His-10 tags in terms of 
crystallization success?

Thanks for your help!
Herman

Re: [ccp4bb] His-6 versus His-10 tag

[Savvas Savvides]

Hi Herman
the survey article by Carson in Acta D DOI: 10.1107/S0907444906052024 
 from a decade ago may provide 
relevant information/insights.
best wishes
Savvas




> On 19 Sep 2017, at 12:10, herman.schreu...@sanofi.com wrote:
> 
> Dear BB,
>  
> We are planning the production of a protein for crystallization. From 
> literature, we know that the construct with a 6-histidine tag crystallizes. 
> However, for other biophysical measurements, we would prefer to have a 
> 10-histidine tag.
>  
> Does anyone has experience with His-6 versus His-10 tags in terms of 
> crystallization success?
>  
> Thanks for your help!
> Herman

Re: [ccp4bb] His-6 versus His-10 tag

[Artem Evdokimov]

Option 1: make two constructs and compare :)

Option 2: put the his10 on the n-term of the protein with a cleavage site

In my experience it matters. I had cases where his4 crystallized and his6
did not. I also had the opposite experience.

Artem


On Sep 19, 2017 6:11 AM,  wrote:

Dear BB,



We are planning the production of a protein for crystallization. From
literature, we know that the construct with a 6-histidine tag crystallizes.
However, for other biophysical measurements, we would prefer to have a
10-histidine tag.



Does anyone has experience with His-6 versus His-10 tags in terms of
crystallization success?



Thanks for your help!

Herman

Re: [ccp4bb] His-6 versus His-10 tag

[Mohinder Pal]

Dear Herman,
I have crystallised the same protein with 6xHis and 12xHis and they both 
diffracted to high resolution. However, I have not seen these tags in the 
structure. 

Best wishes,

Mohinder 

Sent from my iPhone

> On 19 Sep 2017, at 12:10, herman.schreu...@sanofi.com wrote:
> 
> Dear BB,
>  
> We are planning the production of a protein for crystallization. From 
> literature, we know that the construct with a 6-histidine tag crystallizes. 
> However, for other biophysical measurements, we would prefer to have a 
> 10-histidine tag.
>  
> Does anyone has experience with His-6 versus His-10 tags in terms of 
> crystallization success?
>  
> Thanks for your help!
> Herman
>  
>  

Re: [ccp4bb] His-6 versus His-10 tag

[Parthasarathy Sampathkumar]

Hi Herman,

I worked on a kinase, where moved from 6-His in the literature to 8-His,
and it didn't impact crystallization or diffraction (which around 3Angs).

Good Luck
Partha
On Tue, Sep 19, 2017 at 9:43 AM Oganesyan, Vaheh <oganesy...@medimmune.com>
wrote:

> Hi Herman,
>
>
>
> I haven’t done His-6 versus His-10 for the same protein, but have done
> that for different ones with success. However, if in His-6 containing
> protein structure the packing or folding is such that you don’t see His-6
> then it shouldn’t matter it is 6 or 10. Just an opinion.
>
>
>
> *Regards,*
>
>
>
> *Vaheh Oganesyan*
>
> *www.medimmune.com <http://www.medimmune.com>*
>
>
>
> *From:* CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] *On Behalf Of *
> herman.schreu...@sanofi.com
> *Sent:* Tuesday, September 19, 2017 6:11 AM
> *To:* CCP4BB@JISCMAIL.AC.UK
> *Subject:* [ccp4bb] His-6 versus His-10 tag
>
>
>
> Dear BB,
>
>
>
> We are planning the production of a protein for crystallization. From
> literature, we know that the construct with a 6-histidine tag crystallizes.
> However, for other biophysical measurements, we would prefer to have a
> 10-histidine tag.
>
>
>
> Does anyone has experience with His-6 versus His-10 tags in terms of
> crystallization success?
>
>
>
> Thanks for your help!
>
> Herman
>
>
>
>
> To the extent this electronic communication or any of its attachments
> contain information that is not in the public domain, such information is
> considered by MedImmune to be confidential and proprietary. This
> communication is expected to be read and/or used only by the individual(s)
> for whom it is intended. If you have received this electronic communication
> in error, please reply to the sender advising of the error in transmission
> and delete the original message and any accompanying documents from your
> system immediately, without copying, reviewing or otherwise using them for
> any purpose. Thank you for your cooperation.
>

Re: [ccp4bb] His-6 versus His-10 tag

[Oganesyan, Vaheh]

Hi Herman,

I haven't done His-6 versus His-10 for the same protein, but have done that for 
different ones with success. However, if in His-6 containing protein structure 
the packing or folding is such that you don't see His-6 then it shouldn't 
matter it is 6 or 10. Just an opinion.

Regards,

Vaheh Oganesyan
www.medimmune.com

From: CCP4 bulletin board [mailto:CCP4BB@JISCMAIL.AC.UK] On Behalf Of 
herman.schreu...@sanofi.com
Sent: Tuesday, September 19, 2017 6:11 AM
To: CCP4BB@JISCMAIL.AC.UK
Subject: [ccp4bb] His-6 versus His-10 tag

Dear BB,

We are planning the production of a protein for crystallization. From 
literature, we know that the construct with a 6-histidine tag crystallizes. 
However, for other biophysical measurements, we would prefer to have a 
10-histidine tag.

Does anyone has experience with His-6 versus His-10 tags in terms of 
crystallization success?

Thanks for your help!
Herman


To the extent this electronic communication or any of its attachments contain 
information that is not in the public domain, such information is considered by 
MedImmune to be confidential and proprietary. This communication is expected to 
be read and/or used only by the individual(s) for whom it is intended. If you 
have received this electronic communication in error, please reply to the 
sender advising of the error in transmission and delete the original message 
and any accompanying documents from your system immediately, without copying, 
reviewing or otherwise using them for any purpose. Thank you for your 
cooperation.

[ccp4bb] His-6 versus His-10 tag

[Herman . Schreuder]

Dear BB,

We are planning the production of a protein for crystallization. From 
literature, we know that the construct with a 6-histidine tag crystallizes. 
However, for other biophysical measurements, we would prefer to have a 
10-histidine tag.

Does anyone has experience with His-6 versus His-10 tags in terms of 
crystallization success?

Thanks for your help!
Herman