SPring-8, the large synchrotron radiation facility

Inquiry number

SOL-0000001187

Beamline

BL41XU (Macromolecular Crystallography I)

Scientific keywords

A. Sample category	biology, medicine
B. Sample category (detail)	biomolecule, crystal, protein, pharmaceuticals
C. Technique	X-ray diffraction
D. Technique (detail)	single crystal
E. Particular condition	low-T (~ liquid N2)
F. Photon energy	X-ray (4-40 keV)
G. Target information	local structure, structure analysis, crystal structure, morphology

Industrial keywords

level 1---Application area	Pharmaceuticals
level 2---Target	drug design, process analytical technology (PAT)
level 3---Target (detail)	protein, drug
level 4---Obtainable information	film thickness, d-spacing (lattice parameter), crystal structure, supra-molecular assemblies
level 5---Technique	diffraction

Classification

A80.50 Pharmaceuticals, M10.10 single crystal diffraction

Body text

Some protein crystals have a large unit-cell over several hundred angstrom depending on the molecular weight, molecular numbers in an asymmetric unit or space group of crystal. These crystals can diffract X-ray weakly because that diffraction intensity is in inverse proportion to the volume of the crystal. Moreover, it is difficult to separate each diffraction spot on the detector.

In this case, it is introduced that the recording of X-ray diffraction image from the crystal with large unit-cell of 590 Å.

This image was collected from crystal of calcium ATPase from rat sarcoplasmic reticulum. The crystal was soaked to aurothioglucose solution as a contrast medium to visualize the lipid bilayer in it.

Using high brilliant and high parallel beam from undulator of BL41XU, each diffraction spots were separated clearly on the detector surface (Fig. 1).

Figure 1. X-ray diffraction image of calcium ATPase from rat sarcoplasmic reticulum
Space group and unit cell of the crystal is P4₁ and a=b=71.7, c=590.3 Å.
Using wavelength is 1.0401 Å, and the used detector is RIGAKU R-AXIS V with 600 mm distance.

Source of the figure

Bulletin from SPring-8

Bulletin title

C. Toyoshima and T. Tsuda, SPring-8 User Experimental Report 12, 212 (2003B)

Page

212

Technique

Source of the figure

No figure

Required time for experimental setup

8 hour(s)

Instruments

Instrument	Purpose	Performance
Protein Crystal Diffractometer	To record diffraction data
Protein Crystal Diffractometer	To record diffraction image	sensitive area: 400mm x 400mm

References

Document name
C. Toyoshima and T. Tsuda, SPring-8 User Experimental Report 12, 212 (2003B)

Related experimental techniques

Questionnaire

The measurement was possible only in SPring-8. Impossible or very difficult in other facilities.
This solution is an application of a main instrument of the beamline.
With user's own instruments.

Ease of measurement

With a great skill

Ease of analysis

With a great skill

How many shifts were needed for taking whole data in the figure?

Two-three shifts