2CE2: Crystal Structure Analysis Of A Fluorescent Form Of H-ras P21 In Complex With Gdp

Citation:
Abstract
We present a new design for a fluorescence microspectrophotometer for use in kinetic crystallography in combination with x-ray diffraction experiments. The FLUMIX device (Fluorescence spectroscopy to monitor intermediates in x-ray crystallography) is built for 0 degrees fluorescence detection, which has several advantages in comparison to a conventional fluorometer with 90 degrees design. Due to the reduced spatial requirements and the need for only one objective, the system is highly versatile, easy to handle, and can be used for many different applications. In combination with a conventional stereomicroscope, fluorescence measurements or reaction initiation can be performed directly in a hanging drop crystallization setup. The FLUMIX device can be combined with most x-ray sources, normally without the need of a specialized mechanical support. As a biological model system, we have used H-Ras p21 with an artificially introduced photo-labile GTP precursor (caged GTP) and a covalently attached fluorophore (IANBD amide). Using the FLUMIX system, detailed information about the state of photolyzed crystals of the modified H-Ras p21 (p21(mod)) could be obtained. Measurements in combination with a synchrotron beamline showed significant fluorescence changes in p21(mod) crystals even within a few seconds of x-ray exposure at 100 K.
PDB ID: 2CE2Download
MMDB ID: 128824
PDB Deposition Date: 2006/2/2
Updated in MMDB: 2017/06
Experimental Method:
x-ray diffraction
Resolution: 1  Å
Source Organism:
Similar Structures:
Biological Unit for 2CE2: monomeric; determined by author and by software (PISA)
Molecular Components in 2CE2
Label Count Molecule
Protein (1 molecule)
1
Gtpase Hras(Gene symbol: HRAS)
Molecule annotation
Chemicals (3 molecules)
1
1
2
1
3
1
* Click molecule labels to explore molecular sequence information.

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