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There are an number (WT & mutants) of X-ray structures published on
xylose isomerase from A. missouriensis (see e.g. 1XIM).
This is a highly negatively-charged protein with a pI of 3.2-3.5.
Then, there is cytochrome C with pI > 10 (a number of entries in the PDB).
Best luck,
Pr. Nadir T. Mrabet
Cellular & Molecular Biochemistry
INSERM U-724
UHP - Nancy 1, School of Medicine
Avenue de la Foret de Haye, BP 184
54505 Vandoeuvre-les-Nancy Cedex
France
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Kornelius Zeth wrote:
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Dear all,
we are trying to crystallize a protein (40 kD) that was
cloned de novo as construct of five identical repeat
structures. The protein contains no LYS, ARG, GLN, GLU,
CYS, MET but a large number of negative charges (pI 3.7).
The protein appears properly folded from CD, size
exclusion. Crystallization hasn't worked out at 10 - 50
mg/ml. We also tried 10 - 50 mg/ml in presence of Ca and
Zn, with and without Histag, we also added small diamines.
Nothing worked so far.
Recloning and mutational changes are not easy due the
repetitive gene structure, however they are possible as the
last anchor (including gene synthesis).
1. As we are very much interested in the protein structure
I wonder whether people from other labs have any experience
with highly charged proteins?
2. Are there any proteins with a significant surplus of
positive charges crystallized that might be
co-crystallized?
3. Did people use any chemicals to modify ASPs or GLUs side
chains?
Best wishes and thanks for your help!
Kornelius
----------------------------------------------
Kornelius Zeth
Max Planck Institute for Developmental Biology
Dept. Protein Evolution
Spemannstr. 35
72076 Tuebingen, Germany
[EMAIL PROTECTED]
Tel -49 7071 601 323
Fax -49 7071 601 349
