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CCP4BB  February 2007

CCP4BB February 2007

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Subject:

Re: Question about glove boxes for protein crystallization

From:

Patrick Shaw Stewart <[log in to unmask]>

Reply-To:

Patrick Shaw Stewart <[log in to unmask]>

Date:

Fri, 23 Feb 2007 11:01:59 +0200

Content-Type:

text/plain

Parts/Attachments:

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text/plain (65 lines)

Hi Mathews

One of our customers, Bret Dillard at the University of Georgia, has
had great success with one of our robots in a Bactron X anaerobic
chamber.

In this case, Bret mainly used the microbatch-under-oil method, which
works very well for anaerobic work:  (1) the oil protects the sample
and reduces exposure to oxygen, (2) the amount of work is far less
because you can keep several degassed screens in the chamber to use
many times with different protein samples.  With vapor diffusion you
will have to degass the solutions every for every few samples (if not
every sample).

Bret won our competition for this work last year.  You can find his report at
http://www.douglas.co.uk/news.htm, plus more info below

Patrick



_________________________________________

Douglas Instruments has announced the winner of the second round of
its competition for the best new crystallization technique.
Congratulations to Bret Dillard from the University of Georgia, for
his winning entry, "Automatic Protein Crystallization in an Anaerobic
Environment."

Bret placed an Oryx1-6 robot in a Bactron X anaerobic chamber, and
used mainly microbatch-under-oil crystallization to crystallize four
proteins that are not stable in an environment with oxygen.  He found
that microbatch avoided the need for frequent degassing of solutions
which reduced the work-load enormously.  The oil also provided extra
protection from oxidation.

One of the proteins crystallized is rubrerythrin from the
hyperthermophilic archaeon, Pyrococcus furiosus.  The native protein
contains iron, but is unstable in oxygen.  A previously-reported
structure was determined in the presence of oxygen, but the iron had
been replaced by zinc.  Using the anaerobic system, Bret has now
obtained the native form containing iron.

_________________________________________



On 2/23/07, Mathews, Irimpan <[log in to unmask]> wrote:
>
>
> Dear Friends,
>
> Sorry for the non CCP4 question. We are planning to purchase a small glove
> box to setup crystallization trays under anaerobic conditions. If you have
> used glove boxes for crystallization, would you please give me some idea?
>
> We are thinking of getting the 815 series from Plas-labs (link below).
>
> http://www.plas-labs.com/
>
> Thank you very much,
> Mathews
>
> Ps: If others are interested, I will post a summary.
>

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