Re: [ccp4bb] Hybrid enzymes

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CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999

Previous message:
Subject: Sequence of crystallised protein fragment
From: Klaus Futterer k {- dot -} futterer {- at -} BHAM {- dot -} AC {- dot -} UK
Date: 2008-07-01

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Subject: Re: Hybrid enzymes
From: "R {- dot -} M {- dot -} Garavito" garavito {- at -} MSU {- dot -} EDU
Date: 2008-07-01

Subject: Re: Hybrid enzymes
From: fenguita {- at -} ITQB {- dot -} UNL {- dot -} PT fenguita {- at -} ITQB {- dot -} UNL {- dot -} PT
Date: 2008-07-01

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
>

-----------------------------------------
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 : fenguita@itqb.unl.pt
-----------------------------------------

CCP4bb navigation

CCP4bb <-- 1999 <-- November 1999 <-- 30 November 1999

Previous message:
Subject: Sequence of crystallised protein fragment
From: Klaus Futterer k {- dot -} futterer {- at -} BHAM {- dot -} AC {- dot -} UK
Date: 2008-07-01

Next message:
Subject: Re: Hybrid enzymes
From: "R {- dot -} M {- dot -} Garavito" garavito {- at -} MSU {- dot -} EDU
Date: 2008-07-01