| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
|
|
|
|||||||
|
|||||||||
From the
1 From the Department of Biochemistry, Stanford University School of Medicine, Stanford, California 94305
N-(Phosphonacetyl)-l-aspartate (PALA) is a stable analogue of the transition state in the reaction catalyzed by aspartate transcarbamylase from Escherichia coli; it combines in 1 molecule most of the structural features of the two natural substrates, carbamyl phosphate and l-aspartate. The inhibition constant (Ki) for PALA with the catalytic subunit is 2.7 x 10-8m at 28° and pH 7.0 in 0.2 m imidazole acetate; the value of Ki is approximately the same with the native enzyme. PALA binds to aspartate transcarbamylase about 1000 times more tightly than carbamyl phosphate, the substrate bound most tightly. In addition to tight binding, other data indicate that PALA interacts with the enzyme at both the carbamyl phosphate and l-aspartate sites simultaneously and puts the enzyme into the same contracted conformation as do carbamyl phosphate and succinate (an analogue of l-aspartate) acting together. PALA alone or carbamyl phosphate and succinate together produce the same ultraviolet difference spectrum with the catalytic subunit; the binding of PALA alone or carbamyl phosphate and succinate together at some of the catalytic sites in the native enzyme activates the remaining catalytic sites to the same extent. PALA gives competitive inhibition with carbamyl phosphate but not with l-aspartate, indicating an obligatory binding order for the substrates: carbamyl phosphate must bind before l-aspartate for catalysis to occur. Titration of the catalytic subunit with PALA, monitored by ultraviolet difference spectroscopy, indicates that there are 3.07 active sites per 100,000 daltons. The interaction of PALA with the enzyme is consistent with the "compression" model for aspartate transcarbamylase previously proposed (Collins, K.D., and Stark, G.R., J. Biol. Chem., 244, 1869 (1969)).
Aspartate Transcarbamylase
INTERACTION WITH THE TRANSITION STATE ANALOGUE N-(PHOSPHONACETYL)-l-ASPARTATE
CiteULike 
Complore 
Connotea 
Del.icio.us 
Digg 
Reddit 
Technorati What's this?
This article has been cited by other articles:
|
|
 |
|
  J. L. Llacer, L. M. Polo, S. Tavarez, B. Alarcon, R. Hilario, and V. Rubio The Gene Cluster for Agmatine Catabolism of Enterococcus faecalis: Study of Recombinant Putrescine Transcarbamylase and Agmatine Deiminase and a Snapshot of Agmatine Deiminase Catalyzing Its Reaction J. Bacteriol., February 15, 2007; 189(4): 1254 - 1265. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. Fetler, E. R. Kantrowitz, and P. Vachette Direct observation in solution of a preexisting structural equilibrium for a mutant of the allosteric aspartate transcarbamoylase PNAS, January 9, 2007; 104(2): 495 - 500. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 G. R. Stark My Life in Science, Not the Restaurant Business J. Biol. Chem., March 18, 2005; 280(11): 9753 - 9760. [Full Text] [PDF]
|
|
|
|
|
|
N. Alam, K. A. Stieglitz, M. D. Caban, S. Gourinath, H. Tsuruta, and E. R. Kantrowitz 240s Loop Interactions Stabilize the T State of Escherichia coli Aspartate Transcarbamoylase J. Biol. Chem., May 28, 2004; 279(22): 23302 - 23310. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
C. Purcarea, A. Ahuja, T. Lu, L. Kovari, H. I. Guy, and D. R. Evans Aquifex aeolicus Aspartate Transcarbamoylase, an Enzyme Specialized for the Efficient Utilization of Unstable Carbamoyl Phosphate at Elevated Temperature J. Biol. Chem., December 26, 2003; 278(52): 52924 - 52934. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 L. Fetler, P. Tauc, D.P. Baker, C.P. Macol, E.R. Kantrowitz, and P. Vachette Replacement of Asp-162 by Ala prevents the cooperative transition by the substrates while enhancing the effect of the allosteric activator ATP on E. coli aspartate transcarbamoylase Protein Sci., May 1, 2002; 11(5): 1074 - 1081. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K.-B. Lee, D. Wang, S. J. Lippard, and P. A. Sharp Transcription-coupled and DNA damage-dependent ubiquitination of RNA polymerase II in vitro PNAS, April 2, 2002; 99(7): 4239 - 4244. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. T. Beernink, Y. R. Yang, R. Graf, D. S. King, S. S. Shah, and H. K. Schachman Random circular permutation leading to chain disruption within and near {{alpha}} helices in the catalytic chains of aspartate transcarbamoylase: Effects on assembly, stability, and function Protein Sci., March 1, 2001; 10(3): 528 - 537. [Abstract] [Full Text]
|
|
|
|
 |
|
 V. Gottifredi, O. Karni-Schmidt, S.-Y. Shieh, and C. Prives p53 Down-Regulates CHK1 through p21 and the Retinoblastoma Protein Mol. Cell. Biol., February 15, 2001; 21(4): 1066 - 1076. [Abstract] [Full Text]
|
|
|
|
 |
|
 G. Seidita, D. Polizzi, G. Costanzo, S. Costa, and A. Di Leonardo Differential gene expression in p53-mediated G1 arrest of human fibroblasts after {{gamma}}-irradiation or N-phosphoacetyl-L-aspartate treatment Carcinogenesis, December 1, 2000; 21(12): 2203 - 2210. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
Y. Qiu and J. N. Davidson Substitutions in the aspartate transcarbamoylase domain of hamster CAD disrupt oligomeric structure PNAS, January 4, 2000; 97(1): 97 - 102. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
V. Serre, H. Guy, B. Penverne, M. Lux, A. Rotgeri, D. Evans, and G. Herve Half of Saccharomyces cerevisiae Carbamoyl Phosphate Synthetase Produces and Channels Carbamoyl Phosphate to the Fused Aspartate Transcarbamoylase Domain J. Biol. Chem., August 20, 1999; 274(34): 23794 - 23801. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
P. T. Beernink, J. A. Endrizzi, T. Alber, and H. K. Schachman Assessment of the allosteric mechanism of aspartate transcarbamoylase based on the crystalline structure of the unregulated catalytic subunit PNAS, May 11, 1999; 96(10): 5388 - 5393. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
B. P. Burns, G. L. Mendz, and S. L. Hazell A Novel Mechanism for Resistance to the Antimetabolite N-Phosphonoacetyl-L-Aspartate by Helicobacter pylori J. Bacteriol., November 1, 1998; 180(21): 5574 - 5579. [Abstract] [Full Text]
|
|
|
|
|
|
Y. Ha, M. T. McCann, M. Tuchman, and N. M. Allewell Substrate-induced conformational change in a trimeric ornithine transcarbamoylase PNAS, September 2, 1997; 94(18): 9550 - 9555. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K. A. Smith, O. B. Chernova, R. P. Groves, M. B. Stark, J. L. Martinez, J. N. Davidson, J. M. Trent, T. E. Patterson, A. Agarwal, P. Duncan, et al. Multiple mechanisms of N-phosphonacetyl-L-aspartate resistance in human cell lines: Carbamyl-P synthetase/aspartate transcarbamylase/dihydro-orotase gene amplification is frequent only when chromosome 2 is rearranged PNAS, March 4, 1997; 94(5): 1816 - 1821. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. M. Aucoin, E. J. Pishko, D. P. Baker, and E. R. Kantrowitz Engineered Complementation in Escherichia coli Aspartate Transcarbamoylase. HETEROTROPIC REGULATION BY QUATERNARY STRUCTURE STABILIZATION J. Biol. Chem., November 22, 1996; 271(47): 29865 - 29869. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S P Linke, K C Clarkin, A Di Leonardo, A Tsou, and G M Wahl A reversible, p53-dependent G0/G1 cell cycle arrest induced by ribonucleotide depletion in the absence of detectable DNA damage. Genes & Dev., April 15, 1996; 10(8): 934 - 947. [Abstract] [PDF]
|
|
|
|
|
|
K. Wei and B. E. Huber Cytosine Deaminase Gene as a Positive Selection Marker J. Biol. Chem., February 16, 1996; 271(7): 3812 - 3816. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
G. Ricci, M. Lo Bello, A. M. Caccuri, A. Pastore, M. Nuccetelli, M. W. Parker, and G. Federici Site-directed Mutagenesis of Human Glutathione Transferase P1-1 J. Biol. Chem., January 20, 1995; 270(3): 1243 - 1248. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. Kantrowitz and W. Lipscomb Escherichia coli aspartate transcarbamylase: the relation between structure and function Science, August 5, 1988; 241(4866): 669 - 674. [Abstract] [PDF]
|
|
|
|
|
|
D. B. Langley, M. D. Templeton, B. A. Fields, R. E. Mitchell, and C. A. Collyer Mechanism of Inactivation of Ornithine Transcarbamoylase by Ndelta -(N'-Sulfodiaminophosphinyl)-L-ornithine, a True Transition State Analogue?. CRYSTAL STRUCTURE AND IMPLICATIONS FOR CATALYTIC MECHANISM J. Biol. Chem., June 23, 2000; 275(26): 20012 - 20019. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
S. H. Khan, J. Moritsugu, and G. M. Wahl Differential requirement for p19ARF in the p53-dependent arrest induced by DNA damage, microtubule disruption, and ribonucleotide depletion PNAS, March 28, 2000; 97(7): 3266 - 3271. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
K.-B. Lee, D. Wang, S. J. Lippard, and P. A. Sharp Transcription-coupled and DNA damage-dependent ubiquitination of RNA polymerase II in vitro PNAS, April 2, 2002; 99(7): 4239 - 4244. [Abstract] [Full Text] [PDF]
|
|
|
|
|
|
J. A. Endrizzi, P. T. Beernink, T. Alber, and H. K. Schachman Binding of bisubstrate analog promotes large structural changes in the unregulated catalytic trimer of aspartate transcarbamoylase: Implications for allosteric regulation PNAS, May 9, 2000; 97(10): 5077 - 5082. [Abstract] [Full Text] [PDF]
|
|
| HOME | HELP | FEEDBACK | SUBSCRIPTIONS | ARCHIVE | SEARCH | TABLE OF CONTENTS |
| All ASBMB Journals | Molecular and Cellular Proteomics |
| Journal of Lipid Research | ASBMB Today |
