Hello, can you use the pisa server or similar to compare the extent of domain-domain and domain-DNA contacts versus crystal contacts in your structures? It might help to show which is the dominant factor affecting domain orientation. Sent from Yahoo Mail on Android On Tue, 28 May 2019 at 14:45, Zizi Tian<[email protected]> wrote: Hi All,I has recently solved my X-ray structures of the ZnF domains of Bo and its complex with dsDNA. A structural comparison of holo-Bo and the Bo-DNA binary complex reveals that the relative orientation of domains ZnF3 undergoes a rotation of 50.5 degrees Does anyone know how to show there are no effects of the crystal packing . Please help me in this regard. Best regards
At 2019-05-28 20:55:07, "Anamika Singh" <[email protected]> wrote: Hi All, Does anyone know what is the actual difference between CIP and EDTA method for exchanging the GppNHp (non-hydrolysable GTP) or GDP with the purified RAS protein? I have a protocol according to which I need to incubate the purified RAS with GppNHp having 20U of alkaline phosphatase, 10uM of Zinc Chloride and 200mM ammonium sulfate for 2 hr at room temperature. In this protocol, they have mentioned that ammonium sulfate needs to be added in the last. I am quite confused, according to them do I need to add it after 2 hrs incubation or I have to add this at last to the mixture of (purified RAS+GppNHp+Zinc chloride and alkaline phosphatase)? Since this protein and its mechanism is new to me so not sure what to do. After reading some papers also I am not able to understand which method is good for exchange and how these reactions behave? Please help me in this regard. -- Anamika Singh Post-Doctoral FellowSilberman Institute of Life Sciences Hebrew University of Jerusalem, IsraelNo: 054-294-8036 To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1 To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1 ######################################################################## To unsubscribe from the CCP4BB list, click the following link: https://www.jiscmail.ac.uk/cgi-bin/webadmin?SUBED1=CCP4BB&A=1
