The Norwegian Structural Biology Centre (NorStruct) offers three service packages:
SP1:
SP2
SP3:
Protein production
: Structure determination
Drug discovery and design
NorStruct's SP2 “Structure determination” services include protein sample evaluation tasks of quality control and stability/solubility optimization, crystallization trials, data collection, using either the home X-ray source or synchrotron radiation, followed by structure determination and refinement.
Prizes and more details about our standard commissioning service packages can be found at the NorStruct web page http://uit.no/norstruct/ .
Table 1 Standard commissioning in SP2 Structure determination.
Service
Package
Task Specification
(SP)
SP2.1
SP2.2
Quality control
Initial crystallization screening
Dynamic light scattering (DLS) spectra at 4-
PAGE, Mass spectroscopy (MALDI-TOF).
37°C, SDS-
Set-up of 96 robotic nanolitre crystallization conditions at 3
SP2.3
SP2.4
Robotic screening nanolitre crystallization
Buffer optimization different mixing ratios to estimate an optimum protein concentration for subsequent crystallization trials.
Set-up of a minimum of 480 robotic nanolitre crystallization conditions with subsequent grid optimization of promising crystallization conditions identified.
SP2.5 X-ray data collection and structure determination
Search for improved solubility and homogeneity 24 different buffer-conditions (pH-range from 4-9), followed by testing of
12 different additives. Thermofluor methods will be used as a standard.
“Standard” data collection and processing, phasing, using molecular replacement (MR) or single anomalous dispersion
(SAD) techniques, and refinement. More challenging projects can be customized.
The Norwegian Structural Biology Centre
University of Tromsø, N-9037 Tromsø, Telephone: (+47) 77 64 40 00, Fax (+47) 77 64 47 65
Direct telephone: (+47) 77 64 40 70, E-mail: norstruct@chem.uit.no
http://uit.no/norstruct/

Please place tick marks to indicate which tasks are desired. For simplicity this icon
 can be copied and replaced by the empty boxes. Return this electronic form by E-mail to NorStruct, and you will receive in response a tailored contract for placing the final order.
Title ( and NorStruct project number if available a ):
Customer:
Institution:
Adress:
Contact person:
E-mail:
Phone: a Provided by NorStruct
Order:
(Tick  )
SP
SP2.1
Task
Quality control
Specification
SDS-PAGE
Mass spectroscopy:
Date : ………
From the customer
(mandatory)
Estimated purity: ……. %
Provided by the customer
OR
To be done by NorStruct
SP2.2 Initial crystallization screening
SP2.3 Robotic nanolitre crystallization screening
SP2.4 Buffer optimization
.
SP2.5 X-ray data collection and structure determination
The Norwegian Structural Biology Centre
University of Tromsø, N-9037 Tromsø, Telephone (+47) 77 64 40 00, Fax (+47) 77 64 47 65
Direct telephone (+47) 77 64 40 70, E-mail: norstruct@chem.uit.no
http://uit.no/norstruct/
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Details
Protein name:
Batch identification:
Original source:
Information
.
Expression system and vector:
Tag (His, GST, MPB; etc.)
Total amount protein (mg):
Total volume (
 l) b :
Protein concentration (mg/ml):
Indicate Method: OD
280
,
Bradford assay or other
Recommended storage temperature at NorStruct:
- 80 °C
- 20 °C
4°C b When shipping, frozen samples protein should be sent in aliquots of 25-50
NorStruct).
 l (consult
Protein details
Molecular weight (kDa):
Information
Number of amino acids (including the tag): pI (theoretical and/or measured):
Amino acid sequence of this construct:
GenBank accession number:
Please paste the amino acid sequence here (please mark tag and linker regions):
The Norwegian Structural Biology Centre
University of Tromsø, N-9037 Tromsø, Telephone (+47) 77 64 40 00, Fax (+47) 77 64 47 65
Direct telephone (+47) 77 64 40 70, E-mail: norstruct@chem.uit.no
http://uit.no/norstruct/
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Success in crystallization and structure determination requires detailed knowledge of the biochemical and biophysical properties of the protein, including knowledge of all chemicals that may have been carried over from fermentation and purification. Trace chemicals and especially potential ligands may be of particular importance. A brief description of the protocols may be useful to us.
Protein purification steps
Purification steps
Ni 2+ column; ion exchange, gel filtration, etc.
Storage buffer of the protein:
Salt concentration, buffer, pH, additives: etc.
Details
Please list all steps here, including buffers
Stability details pH
Question
1) pH-range where the protein is known to be stable?
2) pH optimum for stability?
Temperature 1) Is the protein particularly unstable at low or high temperatures?
2) Can or must it be frozen at -20 or 80 °C?
3) Temperature optimum for stability?
Additives 1) Are there known buffers or other additives that destabilize the protein?
2) Are there metals or ions that stabilize the protein (Mg 2+ , Zn 2+ , Mn, Cl , SO
4
2, etc)
Stabilizers
Detergents/ salt
1) Should stabilizers, inhibitors, co-factors, etc. be added to the protein solution?
1) Is addition of salt or detergent essential for keeping the sample soluble?
Information
1)
2)
1)
2)
3)
1)
2)
1)
1)
Question Factors
Glycosylation
Cystine/Oxidation
1) Is glycosylation known for this protein sample?
2) Is it at all something to consider?
1) Are there free cysteines in this protein?
2) Can/should we use reducing agents?
3) Should we use oxygen free atmosphere?
Information
1)
2)
3)
The Norwegian Structural Biology Centre
University of Tromsø, N-9037 Tromsø, Telephone (+47) 77 64 40 00, Fax (+47) 77 64 47 65
Direct telephone (+47) 77 64 40 70, E-mail: norstruct@chem.uit.no
http://uit.no/norstruct/
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Polypeptide chains
Membrane
Domains
Oligomerisation
1) Is there more than one polypeptide chain forming the quaternary structure?
1)
1) Is this a membrane or membrane associated protein? Micelles?
1) Are there multiple domains?
2) Are domains separated by flexible hinge regions?
1) Is non-specific oligomerisation a known property?
2) Is the protein a well defined multimer
(dimer, trimer etc.)?
1)
1)
2)
1)
2)
Other details
Please provide NorStruct with contact information in case more information regarding the protein samples, especially with respect to the details of protein purification, is needed:
E-mail Phone Name
The Norwegian Structural Biology Centre
University of Tromsø, N-9037 Tromsø, Telephone (+47) 77 64 40 00, Fax (+47) 77 64 47 65
Direct telephone (+47) 77 64 40 70, E-mail: norstruct@chem.uit.no
http://uit.no/norstruct/
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