1PLJ

CRYSTALLOGRAPHIC STUDIES ON P21H-RAS USING SYNCHROTRON LAUE METHOD: IMPROVEMENT OF CRYSTAL QUALITY AND MONITORING OF THE GTPASE REACTION AT DIFFERENT TIME POINTS

Summary for 1PLJ

• Entry DOI: 10.2210/pdb1plj/pdb
• Descriptor: C-H-RAS P21 PROTEIN, MAGNESIUM ION, GUANOSINE 5'-TRIPHOSPHATE P3-[1-(2-NITROPHENYL)ETHYL ESTER] (3 entities in total)
• Functional Keywords: oncogene protein
• Biological source: Homo sapiens (human)
• Cellular location: Cell membrane; Lipid-anchor; Cytoplasmic side: P01112
• Total number of polymer chains: 1
• Total formula weight: 19611.89

Authors

Scheidig, A.,Sanchez-Llorente, A.,Lautwein, A.,Pai, E.F.,Corrie, J.E.T.,Reid, G.P.,Wittinghofer, A.,Goody, R.S. (deposition date: 1994-03-13, release date: 1994-07-31, Last modification date: 2024-02-14)

Primary citation

Scheidig, A.J.,Sanchez-Llorente, A.,Lautwein, A.,Pai, E.F.,Corrie, J.E.,Reid, G.P.,Wittinghofer, A.,Goody, R.S.
Crystallographic studies on p21(H-ras) using the synchrotron Laue method: improvement of crystal quality and monitoring of the GTPase reaction at different time points.
Acta Crystallogr.,Sect.D, 50:512-520, 1994

PubMed Abstract: The parameters affecting the crystal quality of complexes between p21(H-ras) and caged GTP have been investigated. The use of pure diastereomers of caged GTP complexed to the more stable p21(G12P)' mutant of p21 and the addition of n-octyl-beta-D-glucopyranoside improved the reproducibility and decreased the mosaicity of the crystals significantly. Furthermore, the crystallization technique was changed from the batch method to the sitting-drop technique. With the availability of a larger yield of well ordered crystals, it was possible to extend the time-resolved crystallographic investigations on p21(H-ras). A structure of p21(G12P)':GTP could be obtained 2 min after photolytic removal of the cage group and led to the identification of a previously unidentified conformation for the so-called catalytically active loop L4. The refinement of five data sets collected within 2 min at different times (2-4, 11-13, 20-22, 30-32 and 90-92 min) after the initiation of the intrinsic GTPase reaction of the protein indicates that the synchrotron Laue method can be used to detect small structural changes and alternative conformations, but is presently limited in the analysis of larger rearrangements since these produce diffuse and broken electron density.

PubMed: 15299412
DOI: 10.1107/S090744499301443X

Experimental method

X-RAY DIFFRACTION (2.8 Å)