3cpa: Difference between revisions

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New page: left|200px<br /><applet load="3cpa" size="450" color="white" frame="true" align="right" spinBox="true" caption="3cpa, resolution 2.0Å" /> '''X-RAY CRYSTALLOGRAPHI...
 
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[[Image:3cpa.jpg|left|200px]]<br /><applet load="3cpa" size="450" color="white" frame="true" align="right" spinBox="true"  
[[Image:3cpa.jpg|left|200px]]<br /><applet load="3cpa" size="350" color="white" frame="true" align="right" spinBox="true"  
caption="3cpa, resolution 2.0&Aring;" />
caption="3cpa, resolution 2.0&Aring;" />
'''X-RAY CRYSTALLOGRAPHIC INVESTIGATION OF SUBSTRATE BINDING TO CARBOXYPEPTIDASE A AT SUBZERO TEMPERATURE'''<br />
'''X-RAY CRYSTALLOGRAPHIC INVESTIGATION OF SUBSTRATE BINDING TO CARBOXYPEPTIDASE A AT SUBZERO TEMPERATURE'''<br />


==Overview==
==Overview==
A high-resolution x-ray crystallographic investigation of the complex, between carboxypeptidase A (CPA; peptidyl-L-amino-acid hydrolase, EC, 3.4.17.1) and the slowly hydrolyzed substrate glycyl-L-tyrosine was done, at -9 degrees C. Although this enzyme-substrate complex has been the, subject of earlier crystallographic investigation, a higher resolution, electron-density map of the complex with greater occupancy of the, substrate was desired. All crystal chemistry (i.e., crystal soaking and, x-ray data collection) was performed on a diffractometer-mounted flow, cell, in which the crystal was immobilized. The x-ray data to 1.6-A, resolution have yielded a well-resolved structure in which the zinc ion of, the active site is five-coordinate: three enzyme residues (glutamate-72, histidine-69, and histidine-196) and the carbonyl oxygen and amino, terminus of glycyl-L-tyrosine complete the coordination polyhedron of the, metal. These results confirm that this substrate may be bound in a, nonproductive manner, because the hydrolytically important zinc-bound, water has been displaced and excluded from the active site. It is likely, that all dipeptide substrates of carboxypeptidase A that carry an, unprotected amino terminus are poor substrates because of such favorable, bidentate coordination to the metal ion of the active site.
A high-resolution x-ray crystallographic investigation of the complex between carboxypeptidase A (CPA; peptidyl-L-amino-acid hydrolase, EC 3.4.17.1) and the slowly hydrolyzed substrate glycyl-L-tyrosine was done at -9 degrees C. Although this enzyme-substrate complex has been the subject of earlier crystallographic investigation, a higher resolution electron-density map of the complex with greater occupancy of the substrate was desired. All crystal chemistry (i.e., crystal soaking and x-ray data collection) was performed on a diffractometer-mounted flow cell, in which the crystal was immobilized. The x-ray data to 1.6-A resolution have yielded a well-resolved structure in which the zinc ion of the active site is five-coordinate: three enzyme residues (glutamate-72, histidine-69, and histidine-196) and the carbonyl oxygen and amino terminus of glycyl-L-tyrosine complete the coordination polyhedron of the metal. These results confirm that this substrate may be bound in a nonproductive manner, because the hydrolytically important zinc-bound water has been displaced and excluded from the active site. It is likely that all dipeptide substrates of carboxypeptidase A that carry an unprotected amino terminus are poor substrates because of such favorable bidentate coordination to the metal ion of the active site.


==About this Structure==
==About this Structure==
3CPA is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/ ] with ZN as [http://en.wikipedia.org/wiki/ligand ligand]. This structure superseeds the now removed PDB entry 1CPA. Active as [http://en.wikipedia.org/wiki/Carboxypeptidase_A Carboxypeptidase A], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.4.17.1 3.4.17.1] Full crystallographic information is available from [http://ispc.weizmann.ac.il/oca-bin/ocashort?id=3CPA OCA].  
3CPA is a [http://en.wikipedia.org/wiki/Single_protein Single protein] structure of sequence from [http://en.wikipedia.org/wiki/ ] with <scene name='pdbligand=ZN:'>ZN</scene> as [http://en.wikipedia.org/wiki/ligand ligand]. This structure supersedes the now removed PDB entry 1CPA. Active as [http://en.wikipedia.org/wiki/Carboxypeptidase_A Carboxypeptidase A], with EC number [http://www.brenda-enzymes.info/php/result_flat.php4?ecno=3.4.17.1 3.4.17.1] Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=3CPA OCA].  


==Reference==
==Reference==
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[[Category: Carboxypeptidase A]]
[[Category: Carboxypeptidase A]]
[[Category: Single protein]]
[[Category: Single protein]]
[[Category: Lipscomb, W.N.]]
[[Category: Lipscomb, W N.]]
[[Category: ZN]]
[[Category: ZN]]
[[Category: hydrolase (c-terminal peptidase)]]
[[Category: hydrolase (c-terminal peptidase)]]


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