Dear CPMAS,

Definitely try crystallizing with the MBP - you may be lucky, but at the same time think about other strategies.
The fact that your protein aggregates after MBP cleavage may indicate it is not folded correctly. If this is so, you will need to solve that problem first, for example by trying different expression temperature, system, etc.
Also, remember you may have flexibility between the MBP and your protein, which may inhibit crystallization.

Mark J van Raaij
Dpto de Estructura de Macromoleculas
Centro Nacional de Biotecnologia - CSIC
c/Darwin 3
E-28049 Madrid, Spain
tel. (+34) 91 585 4616
http://www.cnb.csic.es/~mjvanraaij

On 12 Aug 2015, at 00:21, Jim Fairman wrote:

> Your success will vary from target to target, but I would give it the good college try.
>
> We recently published on the target Mcl1 with an MBP fusion which allowed us to create a crystallography platform for structure-based drug design:  http://www.ncbi.nlm.nih.gov/pubmed/25909780
>
> Many of the GPCR structures in LCP include fusion proteins in the form of T4-Lysozyme or BRIL:  http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3375611/
>
> Cheers, Jim
>
> On Tue, Aug 11, 2015 at 3:13 PM, CPMAS Chen <[log in to unmask]> wrote:
> Hi, All CCP4 Members,
>
> According to this paper
>
> Crystal structures of fusion proteins with large-affinity tags, Protein Sci. 2003 Jul;12(7):1313-22.
>
> Crystallization with fusion tag is difficult back then, 2003. what is the current consensus on this topic?
>
> I have a protein behaves well with fusion on, but it is easy to aggregate when the MBP fusion is cleaved. So, I wonder what the current thoughts are about crystallization with fusion tag.
>
> Thanks!
>
>
>
> --
>
> ***************************************************
>
> Charles Chen
>
> Research Associate
>
> University of Pittsburgh School of Medicine
>
> Department of Anesthesiology
>
> ******************************************************
>
>
>
>
>
> --
> Jim Fairman, Ph D.
> Group Leader I - Crystallography
> Beryllium
> Tel: 206-780-8914
> Cell: 240-479-6575
> E-mail: [log in to unmask] [log in to unmask]