Douglas and Ronaldo,
I wanted to put in my two cents worth on both of your queries at the
same time. You should look up glycerol dehydrogenase from the yeast
S. pombe. Sp-GlyDH was solved accidently a few years ago by Anne
Mulichak in our group (PDB 1TA9). We were trying to crystallize
another enzyme that was expressed in S. pombe, but Sp-GlyDH also co-
purifed on a Co-Talon column. A common cleavage product of the
target protein (mammalian hexokinase III) would produce the very
minor band at 45 KDa, which was almost exactly the same size as Sp-
GlyDH. The minor band represented much less than 5% of the total
protein. We easily got crystals which turned out to be Sp-GlyDH.
Thus, Sp-GlyDH at less than 0.5 mg/mL will crystallize in the
presence of ~9-10 mg/mL of another "contaminating" protein. Never
did get crystals of mammalian hexokinase III.
Michael
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R. Michael Garavito, Ph.D. Email: [EMAIL PROTECTED]
Biochemistry & Molecular Biology Office: (517) 355-9724
Michigan State University Lab: (517) 353-9125
East Lansing, MI 48824-1319 FAX: (517) 353-9334
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On Feb 22, 2007, at 3:53 PM, Douglas L. Theobald wrote:
On Feb 22, 2007, at 2:35 PM, Nat Echols wrote:
I take it you're only interested in well-characterized and well-
known proteins?
Actually no -- well-characterized is good, but well-known is
unnecessary.
I have a receiver domain that expresses at >100mg/L and forms
crystals right out of most screens that diffract to atomic/
subatomic resolution, but it's still being functionally
characterized and the system it's a part of is of limited interest
outside of a specific field of microbiology. Experimentally,
though, I can't imagine an easier protein to work with.
On Thu, 22 Feb 2007, Douglas L. Theobald wrote:
Hi all,
I'd like to pick the collective brain of crystallographers on
this list -- what are some of the most easily crystallizable
proteins? I'm especially interested in those that over-express
and diffract well, and in ones that might be less well-known
than, say, lysozyme (but nearly as nice).
Douglas
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Douglas L. Theobald
Department of Biochemistry
Brandeis University
Waltham, MA 02454-9110
[EMAIL PROTECTED]
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