2CL0

CRYSTAL STRUCTURE ANALYSIS OF A FLUORESCENT FORM OF H-RAS P21 IN COMPLEX WITH GppNHp

Summary for 2CL0

• Entry DOI: 10.2210/pdb2cl0/pdb
• Related: 121P 1AA9 1AGP 1BKD 1CLU 1CRP 1CRQ 1CRR 1CTQ 1GNP 1GNQ 1GNR 1HE8 1IAQ 1IOZ 1JAH 1JAI 1K8R 1LF0 1LF5 1LFD 1NVU 1NVV 1NVW 1NVX 1P2S 1P2T 1P2U 1P2V 1PLJ 1PLK 1PLL 1Q21 1QRA 1RVD 1WQ1 1XCM 1XD2 1XJ0 1ZVQ 1ZW6 221P 2C5L 2CE2 2CL6 2CL7 2CLC 2CLD 2GDP 2Q21 421P 4Q21 521P 5P21 621P 6Q21 721P 821P
• Descriptor: GTPASE HRAS, MAGNESIUM ION, 2-AMINO-2-HYDROXYMETHYL-PROPANE-1,3-DIOL, ... (6 entities in total)
• Functional Keywords: nucleotide binding protein, methylation, lipoprotein, gtp-binding, fluorescence, gppnhp, membrane, palmitate, prenylation, proto-oncogene, golgi apparatus, disease mutation, nucleotide-binding, guanine nucleotide binding protein
• Biological source: HOMO SAPIENS (HUMAN)
• Cellular location: Cell membrane; Lipid-anchor; Cytoplasmic side: P01112
• Total number of polymer chains: 1
• Total formula weight: 19858.24

Authors

Klink, B.U.,Goody, R.S.,Scheidig, A.J. (deposition date: 2006-04-25, release date: 2006-05-31, Last modification date: 2024-11-20)

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-, 2006

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: 16698776
DOI: 10.1529/BIOPHYSJ.105.078931

Experimental method

X-RAY DIFFRACTION (1.8 Å)