In trying to reproduce a very nice public structure a cloning mis-hap put a -GS- prior to the C-term 6His tag. The resultant crystals had a 500Ang C dimension and 16 molecules in the au. Even a single amino acid could make all the difference....
David Hargreaves
Associate Principal Scientist
_____________________________________________________________________
AstraZeneca
DECS, CP&SS
Mereside, 50F49, Alderley Park, Cheshire, SK10 4TF
Tel +44 (0)01625 518521 Fax +44 (0) 1625 232693
David.Hargreaves @astrazeneca.com
Please consider the environment before printing this e-mail
--------------------------------------------------------------------------
AstraZeneca UK Limited is a company incorporated in England and Wales with registered number: 03674842 and a registered office at 2 Kingdom Street, London, W2 6BD.
Confidentiality Notice: This message is private and may contain confidential, proprietary and legally privileged information. If you have received this message in error, please notify us and remove it from your system and note that you must not copy, distribute or take any action in reliance on it. Any unauthorised use or disclosure of the contents of this message is not permitted and may be unlawful.
Disclaimer: Email messages may be subject to delays, interception, non-delivery and unauthorised alterations. Therefore, information expressed in this message is not given or endorsed by AstraZeneca UK Limited unless otherwise notified by an authorised representative independent of this message. No contractual relationship is created by this message by any person unless specifically indicated by agreement in writing other than email.
Monitoring: AstraZeneca UK Limited may monitor email traffic data and content for the purposes of the prevention and detection of crime, ensuring the security of our computer systems and checking Compliance with our Code of Conduct and Policies.
-----Original Message-----
From: CCP4 bulletin board [mailto:[log in to unmask]] On Behalf Of weliu
Sent: 27 June 2012 02:07
To: [log in to unmask]
Subject: [ccp4bb] The effect of His-tag location on crystallization
Dear all,
We crystallized a protein and found that crystal quality greatly depended on the location of His-tag. When a His-tag was added at the C-terminus, only crystalline precipitate or spherical quasi crystals were grown. However, when the His-tag was moved to the N-terminus, single crystals were grown under a number of conditions, and the best one diffracted to 1.7 angstrom after optimization. I was wondering if there were published reports describing similar cases.
Thank you in advance
Wei Liu
|