[ccp4bb] hydrophilic protein going to aggregate

Hello Crystallographers, I am trying to express and purify a soluble domain of a membrane protein for crystallization. The amino acid content is as below Ala (A) 12 13.8% Arg (R) 10 11.5% Asn (N) 2 2.3% Asp (D) 8 9.2% Cys (C) 1 1.1% Gln (Q) 1 1.1% Glu (E) 4 4.6% Gly (G) 16 18.4% His (H) 3 3.4%

[ccp4bb] ITC with unfolded proteins

Hello everyone, I have a query for the scientists working on protein-protein interaction. It is known that some proteins exist in unfolded or molten globule state and attain structure on interaction with other folded proteins. Many a times, it is difficult to obtain the structure of these

Re: [ccp4bb] ITC with unfolded proteins

The City College of New York Department of Chemistry, MR-1135 160 Convent Avenue New York, NY 10031 Tel. (212) 650-6070 Original message Date: Fri, 14 Mar 2014 18:07:48 +0530 From: CCP4 bulletin board CCP4BB@JISCMAIL.AC.UK (on behalf of Anita P crystals...@gmail.com) Subject: [ccp4bb

Re: [ccp4bb] off topic: rmsf in simulation

entropy to increase in the bound state, although in your case it seems to be the whole protein! On Dec 9, 2012, at 1:05 PM, anita p wrote: Hi All, I am trying to understand the mechanism of protein-peptide interaction in two complexes (protein-pepA and protein-pepB). While trying to perform some

[ccp4bb] off topic: rmsf in simulation

Hi All, I am trying to understand the mechanism of protein-peptide interaction in two complexes (protein-pepA and protein-pepB). While trying to perform some simulation experiments, I find that the* root mean square fluctuation (RMSF) by residues of protein in the complex is higher than that of

[ccp4bb] off topic: protein peptide binding

Hi All, I wanted some advice regarding mapping out Protein-peptide interaction. The peptide is a 12 mer and the protein is 15kDa. Invivo studies suggest that the peptide is binds the protein and helps in transport. Hence I feel it would perhaps transient binding. I know that I should do ITC or

[ccp4bb] generating symmetry related

Dear All, I have a small molecule structure file coordinates in CSD CIF format, I would like to analysis inter-molecular interaction between them by generating symmetry related nearest neighbor structures. I want to store the coordinates of the generated structures and further analysis it using

[ccp4bb] granular precipitate

Hi All, I have set up initial screen in hanging drop trays with a protein of theoritical pI of 8.5. The protein is in acetate buffer 10mM, KCl 100mM and 2% glycerol pH 5 . In 85-90% of the conditions I see granular precipitate in 1 day. I tried to open the coverslip, and touch few drops, They had

[ccp4bb] native gels

Hi All, Has anyone run a native gel for proteins at pI8 . I want to pour my own native gel. Do I run a discontinuous page or a continuous one?? Please help with regards to the buffer system to be used, and the dye to be used. With regards Rashmi

Re: [ccp4bb] drops swelling

, E.A. (2011) Practical Use of Glycerol in Protein Crystallization. Cryst. Growth Des. 11 :2755–2762. http://pubs.acs.org/doi/abs/**10.1021/cg101364mhttp://pubs.acs.org/doi/abs/10.1021/cg101364m Enrico. On Fri, 09 Sep 2011 13:22:53 +0200, anita p crystals...@gmail.com wrote: Dear

[ccp4bb] drops swelling

Dear Crystallographers, I have set hanging drop trays with 2ul of protein and 2 ul of resorvior solution. I have seen in some cases the drops are swelling. My protein buffer has 15% glycerol in it. This is happening mainly when I have peg 400 or peg MME or MPD or Jeffamine in the buffer condition.

[ccp4bb] Protein aggregation and crystallization

Hi All, I am working on a protein which has a membrane spanning region and as cytosolic domain.I have made various deletion constructs of the protein, so that I can have a crystallizable fragment. There is no homologues mentioned in the pdb for this protein. All of these constructs are purified

Re: [ccp4bb] Protein aggregation and crystallization

detergent from the begining itself. Please do not chop off the membrane part keep it and chop some of the unstructured cytosolic part if you want. all the best. let us know if any of these worked Padayatti On Fri, Aug 26, 2011 at 3:03 AM, anita p crystals...@gmail.com wrote: Hi All, I am working

Re: [ccp4bb] Protein aggregation and crystallization

it somehow, maybe by light scattering or centrifugation Good luck Yury -- *From:* CCP4 bulletin board [CCP4BB@JISCMAIL.AC.UK] on behalf of anita p [ crystals...@gmail.com] *Sent:* Friday, August 26, 2011 3:03 AM *To:* CCP4BB@JISCMAIL.AC.UK *Subject:* [ccp4bb

[ccp4bb] detergents

Hi all, I have been trying to purify cytosolic fraction of membrane protein whose domain boundries are unknown. hence I have made a series of deletion constructs. The expression and purification is not a problem. I get good yields of the proteins. But on a gelfiltration column, they run in the

[ccp4bb] Detergents

Hi, I had set up crystallization with a bicine as buffer and peg 400 as precipitant. I used the detergent DDAO/LDAO as an additive to the crystallization drop (one of the hampton additive screen condition, it says 5% on the vial) I have a clear drop and in the centre there is a shiny precipitate

Re: [ccp4bb] Tev Cleavage issue !!

or may not work out. You can also consider cleaving your protein at lower concentration, in the presence of detergents or polyols, etc. Cheers, Artem On Thu, Apr 7, 2011 at 9:37 PM, anita p crystals...@gmail.com wrote: Hi Crystallographers, I am working of 23 Kda protein with a Nterminal

[ccp4bb] Tev Cleavage issue !!

Hi Crystallographers, I am working of 23 Kda protein with a Nterminal His tag and a TEV cleavage site. I am getting crystals with the his tag and tev site intact, but they dont diffract. *Is it probable that they dont diffract because of the extra his tag and the tev site?* I am trying to

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