2EVW
Crystal structure analysis of a fluorescent form of H-Ras p21 in complex with R-caged GTP
Summary for 2EVW
| Entry DOI | 10.2210/pdb2evw/pdb |
| Descriptor | GTPase HRas, MAGNESIUM ION, GUANOSINE 5'-TRIPHOSPHATE P3-[1-(2-NITROPHENYL)ETHYL ESTER], ... (5 entities in total) |
| Functional Keywords | guanine nucleotide binding protein, fluorescence, caged gtp, signaling protein |
| Biological source | Homo sapiens (human) |
| Cellular location | Cell membrane. Isoform 2: Nucleus: P01112 |
| Total number of polymer chains | 1 |
| Total formula weight | 19789.01 |
| Authors | Klink, B.U. (deposition date: 2005-11-01, release date: 2006-05-30, Last modification date: 2024-10-09) |
| Primary citation | Klink, B.U.,Goody, R.S.,Scheidig, A.J. A newly designed microspectrofluorometer for kinetic studies on protein crystals in combination with x-ray diffraction. Biophys.J., 91:981-992, 2006 Cited by PubMed 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. PubMed: 16698776DOI: 10.1529/biophysj.105.078931 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (1.05 Å) |
Structure validation
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