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New page: left|200px<br /><applet load="1kee" size="450" color="white" frame="true" align="right" spinBox="true" caption="1kee, resolution 2.10Å" /> '''Inactivation of the ...
 
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[[Image:1kee.jpg|left|200px]]<br /><applet load="1kee" size="450" color="white" frame="true" align="right" spinBox="true"  
[[Image:1kee.jpg|left|200px]]<br /><applet load="1kee" size="350" color="white" frame="true" align="right" spinBox="true"  
caption="1kee, resolution 2.10&Aring;" />
caption="1kee, resolution 2.10&Aring;" />
'''Inactivation of the Amidotransferase Activity of Carbamoyl Phosphate Synthetase by the Antibiotic Acivicin'''<br />
'''Inactivation of the Amidotransferase Activity of Carbamoyl Phosphate Synthetase by the Antibiotic Acivicin'''<br />


==Overview==
==Overview==
Carbamoyl phosphate synthetase (CPS) from Escherichia coli catalyzes the, formation of carbamoyl phosphate from 2 mol of ATP, bicarbonate, and, glutamine. CPS was inactivated by the glutamine analog, acivicin. In the, presence of ATP and bicarbonate the second-order rate constant for the, inactivation of the glutamine-dependent activities was 4.0 x 10(4) m(-1), s(-1). In the absence of ATP and bicarbonate the second-order rate, constant for inactivation of CPS was reduced by a factor of 200. The, enzyme was protected against inactivation by the inclusion of glutamine in, the reaction mixture. The ammonia-dependent activities were unaffected by, the incubation of CPS with acivicin. These results are consistent with the, covalent labeling of the glutamine-binding site located within the small, amidotransferase subunit. The binding of ATP and bicarbonate to the large, subunit of CPS must also induce a conformational change within the, amidotransferase domain of the small subunit that enhances the, nucleophilic character of the thiol group required for glutamine, hydrolysis. The acivicin-inhibited enzyme was crystallized, and the, three-dimensional structure was determined by x-ray diffraction, techniques. The thiol group of Cys-269 was covalently attached to the, dihydroisoxazole ring of acivicin with the displacement of a chloride ion.
Carbamoyl phosphate synthetase (CPS) from Escherichia coli catalyzes the formation of carbamoyl phosphate from 2 mol of ATP, bicarbonate, and glutamine. CPS was inactivated by the glutamine analog, acivicin. In the presence of ATP and bicarbonate the second-order rate constant for the inactivation of the glutamine-dependent activities was 4.0 x 10(4) m(-1) s(-1). In the absence of ATP and bicarbonate the second-order rate constant for inactivation of CPS was reduced by a factor of 200. The enzyme was protected against inactivation by the inclusion of glutamine in the reaction mixture. The ammonia-dependent activities were unaffected by the incubation of CPS with acivicin. These results are consistent with the covalent labeling of the glutamine-binding site located within the small amidotransferase subunit. The binding of ATP and bicarbonate to the large subunit of CPS must also induce a conformational change within the amidotransferase domain of the small subunit that enhances the nucleophilic character of the thiol group required for glutamine hydrolysis. The acivicin-inhibited enzyme was crystallized, and the three-dimensional structure was determined by x-ray diffraction techniques. The thiol group of Cys-269 was covalently attached to the dihydroisoxazole ring of acivicin with the displacement of a chloride ion.


==About this Structure==
==About this Structure==
1KEE is a [http://en.wikipedia.org/wiki/Protein_complex Protein complex] structure of sequences from [http://en.wikipedia.org/wiki/Escherichia_coli Escherichia coli] with MN, K, CL, PO4, ADP, ORN and NET as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Carbamoyl-phosphate_synthase_(glutamine-hydrolyzing) Carbamoyl-phosphate synthase (glutamine-hydrolyzing)], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=6.3.5.5 6.3.5.5] Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=1KEE OCA].  
1KEE is a [http://en.wikipedia.org/wiki/Protein_complex Protein complex] structure of sequences from [http://en.wikipedia.org/wiki/Escherichia_coli Escherichia coli] with <scene name='pdbligand=MN:'>MN</scene>, <scene name='pdbligand=K:'>K</scene>, <scene name='pdbligand=CL:'>CL</scene>, <scene name='pdbligand=PO4:'>PO4</scene>, <scene name='pdbligand=ADP:'>ADP</scene>, <scene name='pdbligand=ORN:'>ORN</scene> and <scene name='pdbligand=NET:'>NET</scene> as [http://en.wikipedia.org/wiki/ligands ligands]. Active as [http://en.wikipedia.org/wiki/Carbamoyl-phosphate_synthase_(glutamine-hydrolyzing) Carbamoyl-phosphate synthase (glutamine-hydrolyzing)], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=6.3.5.5 6.3.5.5] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=1KEE OCA].  


==Reference==
==Reference==
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[[Category: Escherichia coli]]
[[Category: Escherichia coli]]
[[Category: Protein complex]]
[[Category: Protein complex]]
[[Category: Holden, H.M.]]
[[Category: Holden, H M.]]
[[Category: Miles, B.W.]]
[[Category: Miles, B W.]]
[[Category: Raushel, F.M.]]
[[Category: Raushel, F M.]]
[[Category: Thoden, J.B.]]
[[Category: Thoden, J B.]]
[[Category: ADP]]
[[Category: ADP]]
[[Category: CL]]
[[Category: CL]]
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[[Category: channeling]]
[[Category: channeling]]


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