2CLC

CRYSTAL STRUCTURE ANALYSIS OF A FLUORESCENT FORM OF H-RAS P21 IN COMPLEX WITH GTP (2)

2CLC の概要

• エントリーDOI: 10.2210/pdb2clc/pdb
• 関連するPDBエントリー: 2CL0 2CL6 2CL7 2CLD
• 分子名称: GTPASE HRAS, MAGNESIUM ION, N,N'-DIMETHYL-N-(ACETYL)-N'-(7-NITROBENZ-2-OXA-1,3-DIAZOL-4-YL)ETHYLENEDIAMINE, ... (5 entities in total)
• 機能のキーワード: nucleotide-binding protein, golgi apparatus, proto-oncogene, r-caged gtp, prenylation, lipoprotein, gtp-binding, palmitate, methylation
• 由来する生物種: HOMO SAPIENS (HUMAN)
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 19664.17

構造登録者

Klink, B.U.,Goody, R.S.,Scheidig, A.J. (登録日: 2006-04-27, 公開日: 2006-05-24, 最終更新日: 2024-11-20)

主引用文献

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

実験手法

X-RAY DIFFRACTION (1.3 Å)