Hi Tony,
as for most things protein, there's a kit (presumably several) you
could try to screen a set of proteases:
https://hamptonresearch.com/product_detail.aspx?cid=30&sid=193&pid=593
('Proti Ace').
Bärbel
Quoting Antonio Ariza <[log in to unmask]>:
> Hi all,
>
>
>
> I have a couple of proteins that are stable as full-length
> constructs, but have refused to crystallise after many attempts with
> different buffers and ligands. I'd like to try in-situ proteolysis
> with these before I start cloning truncated versions of them.
>
>
>
> Have any of you used this method and do you have any tips?
>
>
>
> Cheers,
>
>
>
> Tony
>
>
>
> ------------------------------------------------------
>
> Dr. Antonio Ariza
> University of Oxford
> Sir William Dunn School of Pathology
> South Parks Road
> Oxford
> OX1 3RE
> ________________________________
> From: CCP4 bulletin board [[log in to unmask]] on behalf of
> Eleanor Dodson [[log in to unmask]]
> Sent: 30 October 2015 07:37
> To: [log in to unmask]
> Subject: Re: [ccp4bb] Fo-Fo map creation
>
> Well - it is easy in the GUI
> Use cad (reflection utils) to get an mtz file with F_LIg .. F_Papo--
> PHICapo FOM_apo
>
> Then scale it to make sure the Fapo and FLig are on the same scale.
> The analysis of Riso gives some idea of isomorphism
>
> The Map utilities
>
> F1 = FLig F2 = F_apo PHI = PHIC W = FOM
>
> That will give a quick picture, similar to that generated by DIMPLE
> - there are more sophisticated procedures but this should be enough
> to check if the Ligand IS there.
>
> Eleanor
>
> On 29 October 2015 at 19:55, Hena Dutta
> <[log in to unmask]<mailto:[log in to unmask]>> wrote:
> Dear CCP4BB Members,
> I am trying to create Fo-Fo map from two data sets.
> 1. FP_apo, SIGFP_apo
> cell: 160.91, 70.73, 110.51, 90, 130.65, 90; space gr. C2
> &
> 2. F_LigBound, SIGF_LigBound
> cell: 161.03, 69.74, 112.59, 90, 130.31, 90; space gr C2
>
> I have good solution for the receptor with both data sets using PHASER MR.
>
> Phenix could not work as the cell dimensions are not close enough.
> Changing some criteria in phenix may work. But, I don't know how to
> do that.
>
> Can someone tell me the steps in CCP4?
>
> I don't have IMEAN in my mtz file, so could not use DIMPLE. I did
> not find a way to create IMEAN from F in CCP4. Sorry, if my question
> is trivial.
>
> Best regards,
> Hena
--
Bärbel Blaum, Ph.D.
Interfakultäres Institut für Biochemie (IFIB)
Hoppe-Seyler-Strasse 4
D-72076 Tübingen
Germany
+49 70 71 29 75 359
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