Evidence for distinct mechanisms of transition state stabilization of GTPases by fluoride

S Vincent; M Brouns; M J Hart; J Settleman

doi:10.1073/pnas.95.5.2210

Evidence for distinct mechanisms of transition state stabilization of GTPases by fluoride

S Vincent, M Brouns, M J Hart, J Settleman

Research output: Contribution to Journal › Article › Academic › peer-review

Abstract

GTPase-activating proteins (GAPs) function by stabilizing the GTPase transition state. This has been most clearly demonstrated by the formation of a high-affinity complex between various GAPs and GDP-bound GTPases in the presence of aluminum tetrafluoride, which can mimic the gamma-phosphate of GTP. Herein, we report that p190 RhoGAP forms a high-affinity complex with Rho GTPases in the presence of fluoride ions, suggesting that p190 also functions to stabilize the GTPase transition state. However, this Rho-p190 complex does not require aluminum ions or even guanine nucleotide, indicating a distinct role for fluoride that is not consistent with the gamma-phosphate-mimicking hypothesis. These results indicate that it is necessary to reconsider the assumed role of fluoride in stabilizing a variety of other GTPase-GAP interactions where the requirement for aluminum or guanine nucleotide has not yet been addressed.

Original language	English
Pages (from-to)	2210-2215
Number of pages	6
Journal	Proceedings of the National Academy of Sciences of the United States of America
Volume	95
Issue number	5
DOIs	https://doi.org/10.1073/pnas.95.5.2210 https://doi.org/10.1073/pnas.95.5.2210
Publication status	Published - 3 Mar 1998
Externally published	Yes

Keywords

3T3 Cells
Animals
COS Cells
Cell Line
Enzyme Stability
Fibroblasts
GTP Phosphohydrolases/biosynthesis
GTP-Binding Proteins/biosynthesis
GTPase-Activating Proteins
Glutathione Transferase/biosynthesis
Guanosine 5'-O-(3-Thiotriphosphate)/pharmacology
Guanosine Diphosphate/pharmacology
Humans
Kinetics
Mice
Protein Biosynthesis
Proteins/chemistry
Recombinant Fusion Proteins/biosynthesis
Sequence Deletion
Sodium Fluoride/pharmacology
Transfection

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title = "Evidence for distinct mechanisms of transition state stabilization of GTPases by fluoride",

abstract = "GTPase-activating proteins (GAPs) function by stabilizing the GTPase transition state. This has been most clearly demonstrated by the formation of a high-affinity complex between various GAPs and GDP-bound GTPases in the presence of aluminum tetrafluoride, which can mimic the gamma-phosphate of GTP. Herein, we report that p190 RhoGAP forms a high-affinity complex with Rho GTPases in the presence of fluoride ions, suggesting that p190 also functions to stabilize the GTPase transition state. However, this Rho-p190 complex does not require aluminum ions or even guanine nucleotide, indicating a distinct role for fluoride that is not consistent with the gamma-phosphate-mimicking hypothesis. These results indicate that it is necessary to reconsider the assumed role of fluoride in stabilizing a variety of other GTPase-GAP interactions where the requirement for aluminum or guanine nucleotide has not yet been addressed.",

keywords = "3T3 Cells, Animals, COS Cells, Cell Line, Enzyme Stability, Fibroblasts, GTP Phosphohydrolases/biosynthesis, GTP-Binding Proteins/biosynthesis, GTPase-Activating Proteins, Glutathione Transferase/biosynthesis, Guanosine 5'-O-(3-Thiotriphosphate)/pharmacology, Guanosine Diphosphate/pharmacology, Humans, Kinetics, Mice, Protein Biosynthesis, Proteins/chemistry, Recombinant Fusion Proteins/biosynthesis, Sequence Deletion, Sodium Fluoride/pharmacology, Transfection",

author = "S Vincent and M Brouns and Hart, {M J} and J Settleman",

year = "1998",

month = mar,

day = "3",

doi = "10.1073/pnas.95.5.2210",

language = "English",

volume = "95",

pages = "2210--2215",

journal = "Proceedings of the National Academy of Sciences of the United States of America",

issn = "0027-8424",

publisher = "National Acad Sciences",

number = "5",

}

TY - JOUR

T1 - Evidence for distinct mechanisms of transition state stabilization of GTPases by fluoride

AU - Vincent, S

AU - Brouns, M

AU - Hart, M J

AU - Settleman, J

PY - 1998/3/3

Y1 - 1998/3/3

N2 - GTPase-activating proteins (GAPs) function by stabilizing the GTPase transition state. This has been most clearly demonstrated by the formation of a high-affinity complex between various GAPs and GDP-bound GTPases in the presence of aluminum tetrafluoride, which can mimic the gamma-phosphate of GTP. Herein, we report that p190 RhoGAP forms a high-affinity complex with Rho GTPases in the presence of fluoride ions, suggesting that p190 also functions to stabilize the GTPase transition state. However, this Rho-p190 complex does not require aluminum ions or even guanine nucleotide, indicating a distinct role for fluoride that is not consistent with the gamma-phosphate-mimicking hypothesis. These results indicate that it is necessary to reconsider the assumed role of fluoride in stabilizing a variety of other GTPase-GAP interactions where the requirement for aluminum or guanine nucleotide has not yet been addressed.

AB - GTPase-activating proteins (GAPs) function by stabilizing the GTPase transition state. This has been most clearly demonstrated by the formation of a high-affinity complex between various GAPs and GDP-bound GTPases in the presence of aluminum tetrafluoride, which can mimic the gamma-phosphate of GTP. Herein, we report that p190 RhoGAP forms a high-affinity complex with Rho GTPases in the presence of fluoride ions, suggesting that p190 also functions to stabilize the GTPase transition state. However, this Rho-p190 complex does not require aluminum ions or even guanine nucleotide, indicating a distinct role for fluoride that is not consistent with the gamma-phosphate-mimicking hypothesis. These results indicate that it is necessary to reconsider the assumed role of fluoride in stabilizing a variety of other GTPase-GAP interactions where the requirement for aluminum or guanine nucleotide has not yet been addressed.

KW - 3T3 Cells

KW - Animals

KW - COS Cells

KW - Cell Line

KW - Enzyme Stability

KW - Fibroblasts

KW - GTP Phosphohydrolases/biosynthesis

KW - GTP-Binding Proteins/biosynthesis

KW - GTPase-Activating Proteins

KW - Glutathione Transferase/biosynthesis

KW - Guanosine 5'-O-(3-Thiotriphosphate)/pharmacology

KW - Guanosine Diphosphate/pharmacology

KW - Humans

KW - Kinetics

KW - Mice

KW - Protein Biosynthesis

KW - Proteins/chemistry

KW - Recombinant Fusion Proteins/biosynthesis

KW - Sequence Deletion

KW - Sodium Fluoride/pharmacology

KW - Transfection

U2 - 10.1073/pnas.95.5.2210

DO - 10.1073/pnas.95.5.2210

M3 - Article

C2 - 9482864

SN - 0027-8424

VL - 95

SP - 2210

EP - 2215

JO - Proceedings of the National Academy of Sciences of the United States of America

JF - Proceedings of the National Academy of Sciences of the United States of America

IS - 5

ER -

Evidence for distinct mechanisms of transition state stabilization of GTPases by fluoride

Abstract

Keywords

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