Vanadate catalyzes photocleavage of adenylate kinase at proline-17 in the phosphate-binding loop

Biochemistry ◽  
1992 ◽  
Vol 31 (2) ◽  
pp. 491-497 ◽  
Author(s):  
Christine R. Cremo ◽  
Joseph A. Loo ◽  
Charles G. Edmonds ◽  
Kristina M. Hatlelid
Keyword(s):  
Adenylate Kinase ◽  
Phosphate Binding ◽  
Binding Loop

Structurally and catalytically important residues in the phosphate binding loop of adenylate kinase of Escherichia coli

Biochemistry ◽  
1990 ◽  
Vol 29 (32) ◽  
pp. 7451-7459 ◽  
Author(s):  
Jochen Reinstein ◽  
Ilme Schlichting ◽  
Alfred Wittinghofer
Keyword(s):  
Escherichia Coli ◽  
Adenylate Kinase ◽  
Phosphate Binding ◽  
Binding Loop

scholarly journals High Resolution Crystal Structures of Human Rab5a and Five Mutants with Substitutions in the Catalytically Important Phosphate-binding Loop

2002 ◽  
Vol 278 (4) ◽  
pp. 2452-2460 ◽  
Author(s):  
Guangyu Zhu ◽  
Jian Liu ◽  
Simon Terzyan ◽  
Peng Zhai ◽  
Guangpu Li ◽  
...  
Keyword(s):  
High Resolution ◽  
Crystal Structures ◽  
Phosphate Binding ◽  
Binding Loop

scholarly journals Phosphate-binding loop and Rab GTPase function: mutations at Ser29 and Ala30 of Rab5 lead to loss-of-function as well as gain-of-function phenotype

2001 ◽  
Vol 355 (3) ◽  
pp. 681-689 ◽  
Author(s):  
Guangpu LI ◽  
Zhimin LIANG
Keyword(s):  
Structure Function ◽  
Binding Domain ◽  
Loss Of Function ◽  
Phosphate Binding ◽  
Gain Of Function ◽  
Gtp Binding ◽  
Early Endosomes ◽  
Structure Function Relationship ◽  
Function Relationship ◽  
Binding Loop

Ras-like GTPases contain a structurally conserved GTP-binding domain. An important element of the GTP-binding domain is the phosphate-binding loop, which contains two Gly residues (Gly12 and Gly13) in Ras. Because the two Gly residues are crucial for normal Ras function, it is intriguing that they are not conserved in other Ras-like GTPases, including the Rab GTPases; for example, the equivalent residues in Rab5 are Ser29 and Ala30. The present study builds on earlier biochemical characterizations of the Rab5 mutants containing substitutions at Ala30 and provides a comprehensive analysis of the structure–function relationship of the Rab5 phosphate-binding loop. We have generated 19 new mutants containing amino acid substitutions at Ser29 and determined whether these Ser29 mutants, as well as the Ala30 mutants, remain able to stimulate the endocytosis of horseradish peroxidase in baby hamster kidney cells. A total of 11 mutants lose the activity of stimulating endocytosis. Of these 11 mutants, 9 are defective in membrane association. In contrast, 27 mutants remain able to stimulate endocytosis. Five of them induce a novel cellular phenotype: cell rounding and detachment from culture dishes. They also induce super-large early endosomes such as the constitutively activated Rab5:Q79L mutant. Biochemical results suggest that the constitutive activation of Rab5 requires an increased nucleotide exchange rate and/or decreased GTPase activity. This study establishes functional significance for the phosphate-binding loop of Rab5 and shows that mutations in this region lead to either a loss-of-function or a gain-of-function phenotype, indicating a structure–function relationship distinct from that of Ras.

scholarly journals GTPase mechanism and function: new insights from systematic mutational analysis of the phosphate-binding loop residue Ala30 of Rab5

2000 ◽  
Vol 346 (2) ◽  
pp. 501-508 ◽  
Author(s):  
Zhimin LIANG ◽  
Timothy MATHER ◽  
Guangpu LI
Keyword(s):  
Biological Activity ◽  
Mutational Analysis ◽  
The Other ◽  
Hydrolysis Reaction ◽  
Rab Gtpase ◽  
Gtpase Activity ◽  
Phosphate Binding ◽  
Gtp Hydrolysis ◽  
Arginine Residues ◽  
Binding Loop

Structural and biochemical data indicate the importance of the phosphate-binding loop residues Gly12 and Gly13 of Ras both in the GTP hydrolysis reaction and in biological activity, but these two residues are not conserved in other Ras-related GTPases. To gain a better understanding of this region in GTP hydrolysis and GTPase function, we used the Ras-related Rab5 GTPase as a model for comparison, and substituted the Ala30 residue (the equivalent of Gly13 of Ras) with all the other 19 amino acids. The resulting mutants were analysed for GTP hydrolysis, GTP binding, GTP dissociation and biological activity. Only the substitution of alanine with proline reduced the GTPase activity by an order of magnitude. This effect is in sharp contrast with the observation that a proline substitution at the neighbouring position (Gly12 of Ras) has little effect on the GTPase activity. Whereas most other substitutions showed either a small negative effect or no effect on the GTPase activity, the arginine substitution surprisingly stimulated the GTPase activity by 5-fold. Molecular modelling suggests that this built-in arginine mimics the catalytic arginine residues found in trimeric GTPases and GTPase-activating proteins in providing the positive charge to facilitate the GTP hydrolysis reaction. We investigated further the biological activity of the Rab5 mutants in relation to stimulating endocytosis. When expressed in cultured baby hamster kidney cells, both arginine and proline mutants, like wild-type Rab5, stimulated endocytosis. However, the arginine mutant was a more potent stimulator than the proline mutant (3-fold stimulation as against 1.7-fold). The tryptophan mutant, on the other hand, was completely deficient in activity in terms of the stimulation of endocytosis, demonstrating the importance of the phosphate-binding loop in Rab GTPase function.

scholarly journals Mutational Analysis of the Phosphate-Binding Loop of Rhizobium meliloti DctD, a ς54-Dependent Activator

1998 ◽  
Vol 180 (10) ◽  
pp. 2792-2795 ◽  
Author(s):  
Yan Gao ◽  
Ying-Kai Wang ◽  
Timothy R. Hoover
Keyword(s):  
Dna Binding ◽  
Transcriptional Activation ◽  
Atp Hydrolysis ◽  
Mutational Analysis ◽  
Rhizobium Meliloti ◽  
Atp Binding ◽  
Phosphate Binding ◽  
Binding Loop

ABSTRACT The phosphate-binding loop of ς54-dependent activators is thought to participate in ATP binding and/or hydrolysis. Alanine substitutions at positions 3, 4, 6, 7, and 8 of this motif inRhizobium meliloti DctD disrupted transcriptional activation and ATP hydrolysis. Interestingly, substitution of alanine at position 7 also affected DNA binding.

Elucidating Microscopic Events Driven by GTP Hydrolysis Reaction in Ras-GAP System with Semi-reactive Molecular Dynamics Simulation: Alternative Role of Phosphate Binding Loop as Mechanical Energy Storage

2021 ◽  
Author(s):  
Ikuo Kurisaki ◽  
Shigenori Tanaka
Keyword(s):  
Molecular Dynamics ◽  
Energy Storage ◽  
Molecular Dynamics Simulation ◽  
Mechanical Energy ◽  
Dynamics Simulation ◽  
Phosphate Binding ◽  
Gtp Hydrolysis ◽  
Reactive Molecular Dynamics ◽  
Binding Loop

ATPase and GTPase have been widely found as chemical energy-mechanical work transducers, whereas the physicochemical mechanisms are not satisfactorily understood. We addressed the problem by examining John Ross’ conjecture that...

scholarly journals A Transient Interaction between the Phosphate Binding Loop and Switch I Contributes to the Allosteric Network between Receptor and Nucleotide in Gαi1

2014 ◽  
Vol 289 (16) ◽  
pp. 11331-11341 ◽  
Author(s):  
Tarjani M. Thaker ◽  
Maruf Sarwar ◽  
Anita M. Preininger ◽  
Heidi E. Hamm ◽  
T. M. Iverson
Keyword(s):  
Phosphate Binding ◽  
Transient Interaction ◽  
Binding Loop
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