Hi :
Very interesting question that you are pointing out !. I have no
experience with enzyme chimeras, however I think that if you got crystals
and they diffract you should be able to solve the structure by MR using
one or both structures as a seeding model within the chimera.
There is a very interesting review by Smyth and coworkers about the use of
fusion proteins in affinity tags for solving protein structures. I know
that is not your case, but is very similar. Take a look at :
Smyth et al. "Crystal structures of fusion proteins with large-affinity
tags". Protein Sci. 2003 Jul;12(7):1313-22. Review.
Hope you find it interesting
Cheers
Francisco
> Dear All:
> I am working with two enzymes (40 kDa each). The crystal structures of
> the two enzymes are known.
> I have made a hybrid enzyme of the two individual ones.
> This chimeric/hybrid enzyme is soluble and exhibits activity of the two
> individual enzymes.
> My questions are as follows:
> 1. Could I use one of the enzyme's structure to solve the crystal
> structure of the hybrid by MR?
> 2. Has anyone working with such hybrids (?) noticed any increase in
> specific activity of either of the enzymes?
> Interestingly, the two enzymes work on the same substrate polymer. One
> is a hydrolase another is an esterase.
>
> Thanks a lot.
> Subbu
>
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Francisco J. Enguita, Ph.D.
Host-pathogen Interactions Group
Macromolecular Crystallography Laboratory
ITQB
EAN, Av. da República
2781-901 Oeiras
Portugal
Phone : +351-21-4469663
Fax : +351-21-4433644
E-mail : [log in to unmask]
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