5ax8: Difference between revisions
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==Recombinant expression, purification and preliminary crystallographic studies of the mature form of human mitochondrial aspartate aminotransferase== | |||
<StructureSection load='5ax8' size='340' side='right' caption='[[5ax8]], [[Resolution|resolution]] 2.99Å' scene=''> | |||
== Structural highlights == | |||
<table><tr><td colspan='2'>[[5ax8]] is a 4 chain structure. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=5AX8 OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5AX8 FirstGlance]. <br> | |||
</td></tr><tr id='related'><td class="sblockLbl"><b>[[Related_structure|Related:]]</b></td><td class="sblockDat">[[3pdb|3pdb]]</td></tr> | |||
<tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=5ax8 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=5ax8 OCA], [http://pdbe.org/5ax8 PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=5ax8 RCSB], [http://www.ebi.ac.uk/pdbsum/5ax8 PDBsum]</span></td></tr> | |||
</table> | |||
== Function == | |||
[[http://www.uniprot.org/uniprot/AATM_HUMAN AATM_HUMAN]] Catalyzes the irreversible transamination of the L-tryptophan metabolite L-kynurenine to form kynurenic acid (KA). Plays a key role in amino acid metabolism. Important for metabolite exchange between mitochondria and cytosol. Facilitates cellular uptake of long-chain free fatty acids.<ref>PMID:9537447</ref> | |||
<div style="background-color:#fffaf0;"> | |||
== Publication Abstract from PubMed == | |||
Mitochondrial aspartate aminotransferase (mAspAT) was recognized as a moonlighting enzyme because it has not only aminotransferase activity but also a high-affinity long-chain fatty acids (LCFA) binding site. This enzyme plays a key role in amino acid metabolism, biosynthesis of kynurenic acid and transport of the LCFA. Therefore, it is important to study the structure-function relationships of human mAspAT protein. In this work, the mature form of human mAspAT was expressed to a high level in Escherichia coli periplasmic space using pET-22b vector, purified by a combination of immobilized metal-affinity chromatography and cation exchange chromatography. Optimal activity of the enzyme occurred at a temperature of 47.5 masculineC and a pH of 8.5. Crystals of human mAspAT were grown using the hanging-drop vapour diffusion method at 277K with 0.1 M HEPES pH 6.8 and 25%(v/v) Jeffamine((R)) ED-2001 pH 6.8. The crystals diffracted to 2.99 A and belonged to the space group P1 with the unit-cell parameters a =56.7, b = 76.1, c = 94.2 A, alpha =78.0, beta =85.6, gamma = 78.4 masculine. Elucidation of mAspAT structure can provide a molecular basis towards understanding catalysis mechanism and substrate binding site of enzyme. | |||
Recombinant expression, purification and crystallographic studies of the mature form of human mitochondrial aspartate aminotransferase.,Jiang X, Wang J, Chang H, Zhou Y Biosci Trends. 2016 Mar 10;10(1):79-84. doi: 10.5582/bst.2015.01150. Epub 2016, Feb 22. PMID:26902786<ref>PMID:26902786</ref> | |||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |||
</div> | |||
<div class="pdbe-citations 5ax8" style="background-color:#fffaf0;"></div> | |||
== References == | |||
<references/> | |||
__TOC__ | |||
</StructureSection> | |||
[[Category: Chang, H]] | [[Category: Chang, H]] | ||
[[Category: Jiang, X]] | |||
[[Category: Wang, J]] | [[Category: Wang, J]] | ||
[[Category: Zhou, Y]] | [[Category: Zhou, Y]] | ||
[[Category: | [[Category: Aspartate aminotransferase]] | ||
[[Category: Kynurenine aminotransferase-iv]] | |||
[[Category: Plasma membrane fatty acid binding protein]] | |||
[[Category: Three-dimensional structure]] | |||
[[Category: Transferase]] | |||
Revision as of 16:16, 10 May 2016
Recombinant expression, purification and preliminary crystallographic studies of the mature form of human mitochondrial aspartate aminotransferase
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