2005
DOI: 10.1074/jbc.m502240200
|View full text |Cite
|
Sign up to set email alerts
|

Structure of the G60A Mutant of Ras

Abstract: Substituting alanine for glycine at position 60 in v-HRas generated a dominant negative mutant that completely abolished the ability of v-H-Ras to transform NIH 3T3 cells and to induce germinal vesicle breakdown in Xenopus oocytes. The crystal structure of the GppNpbound form of RasG60A unexpectedly shows that the switch regions adopt an open conformation reminiscent of the structure of the nucleotide-free form of Ras in complex with Sos. Critical residues that normally stabilize the guanine nucleotide and the… Show more

Help me understand this report

Search citation statements

Order By: Relevance

Paper Sections

Select...
1
1
1

Citation Types

3
55
0

Year Published

2008
2008
2015
2015

Publication Types

Select...
8

Relationship

0
8

Authors

Journals

citations
Cited by 69 publications
(58 citation statements)
references
References 38 publications
3
55
0
Order By: Relevance
“…Then, the various conformations are screened out by various binding partners among the conformational ensembles to fulfill the biological functions, whose binding sites undergo the corresponding changes to fit these partners. For the activated Ras protein with GTP binding, the NMR structures (95) without the corresponding protein partner did not present the low-frequency transient pocket (the potential intermediate bound state (87)); this experimental information further confirms this theory. So in order to sample the low-frequency neighboring states similar to the intermediate, maintaining this open conformation of the loop will largely reproduce the bound state.…”
Section: Targeting the Kinetic Pockets On The Ras Protein For Lead Gesupporting
confidence: 73%
“…Then, the various conformations are screened out by various binding partners among the conformational ensembles to fulfill the biological functions, whose binding sites undergo the corresponding changes to fit these partners. For the activated Ras protein with GTP binding, the NMR structures (95) without the corresponding protein partner did not present the low-frequency transient pocket (the potential intermediate bound state (87)); this experimental information further confirms this theory. So in order to sample the low-frequency neighboring states similar to the intermediate, maintaining this open conformation of the loop will largely reproduce the bound state.…”
Section: Targeting the Kinetic Pockets On The Ras Protein For Lead Gesupporting
confidence: 73%
“…The results from mutations in switch II were thus unexpected ( Table 2). We mutated all possible residues of switch II to Ala, except Gly-60, which is necessary for the conformational change on GTP binding (49), and Gln-61, which is involved in the GTPase mechanism and is already mutated to Leu in all proteins used in these experiments to stabilize the GTP-bound form. Residues 62-70 in Cdc42 were mutated to alanine, but none significantly affected IQGAP1 binding (Table 2).…”
Section: Resultsmentioning
confidence: 99%
“…We thought this mutant might represent the intrinsic property of the state 1 structure of RasWT because it has a conservative substitution in Thr-35, the residue whose interaction with the ␥-phosphate constitutes a key structural basis for the state transition. Moreover, the GppNHp-bound form of this mutant had been shown by 31 P NMR to predominantly adopt state 1 (10). A past attempt to solve the crystal structure of H-RasT35S-GppNHp had failed to identify the electron density of both the switch I and switch II regions (10).…”
Section: Two Distinct State 1 Crystal Structures Of H-rast35s-gppnhp mentioning
confidence: 99%
“…Moreover, determination of the two distinct crystal structures of H-RasT35S-GppNHp, showing prominent polysterism in the switch I and switch II regions, reveals a pivotal role of the guanine nucleotide-mediated interaction between the two switch regions and its rearrangement by a nucleotide positional change in the state 2 to state 1 transition. Furthermore, the 31 P NMR spectra and crystal structures of the GppNHp-bound forms of M-Ras mutants, carrying various H-Ras-type amino acid substitutions, also reveal the existence of a surface pocket in state 1 and support a similar mechanism based on the nucleotide-mediated interaction and its rearrangement in the state 1 to state 2 transition. Intriguingly, the conformational changes accompanying the state transition mimic those that occurred upon GDP/GTP exchange, indicating a common mechanistic basis inherent in the high flexibility of the switch regions.…”
mentioning
confidence: 99%
See 1 more Smart Citation