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| | <StructureSection load='6a6u' size='340' side='right'caption='[[6a6u]], [[Resolution|resolution]] 1.94Å' scene=''> | | <StructureSection load='6a6u' size='340' side='right'caption='[[6a6u]], [[Resolution|resolution]] 1.94Å' scene=''> |
| | == Structural highlights == | | == Structural highlights == |
| - | <table><tr><td colspan='2'>[[6a6u]] is a 1 chain structure with sequence from [http://en.wikipedia.org/wiki/A._nidulans A. nidulans]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=6A6U OCA]. For a <b>guided tour on the structure components</b> use [http://oca.weizmann.ac.il/oca-docs/fgij/fg.htm?mol=6A6U FirstGlance]. <br> | + | <table><tr><td colspan='2'>[[6a6u]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Aspergillus_nidulans Aspergillus nidulans]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=6A6U OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=6A6U FirstGlance]. <br> |
| - | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat"><scene name='pdbligand=D1D:(4S,5S)-1,2-DITHIANE-4,5-DIOL'>D1D</scene>, <scene name='pdbligand=FAD:FLAVIN-ADENINE+DINUCLEOTIDE'>FAD</scene>, <scene name='pdbligand=FSA:1-S-(CARBOXYMETHYL)-1-THIO-BETA-D-FRUCTOPYRANOSE'>FSA</scene>, <scene name='pdbligand=SO4:SULFATE+ION'>SO4</scene></td></tr> | + | </td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 1.945Å</td></tr> |
| - | <tr id='NonStdRes'><td class="sblockLbl"><b>[[Non-Standard_Residue|NonStd Res:]]</b></td><td class="sblockDat"><scene name='pdbligand=CSO:S-HYDROXYCYSTEINE'>CSO</scene></td></tr>
| + | <tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=CSO:S-HYDROXYCYSTEINE'>CSO</scene>, <scene name='pdbligand=D1D:(4S,5S)-1,2-DITHIANE-4,5-DIOL'>D1D</scene>, <scene name='pdbligand=FAD:FLAVIN-ADENINE+DINUCLEOTIDE'>FAD</scene>, <scene name='pdbligand=FSA:1-S-(CARBOXYMETHYL)-1-THIO-BETA-D-FRUCTOPYRANOSE'>FSA</scene>, <scene name='pdbligand=SO4:SULFATE+ION'>SO4</scene></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=6a6u FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=6a6u OCA], [http://pdbe.org/6a6u PDBe], [http://www.rcsb.org/pdb/explore.do?structureId=6a6u RCSB], [http://www.ebi.ac.uk/pdbsum/6a6u PDBsum], [http://prosat.h-its.org/prosat/prosatexe?pdbcode=6a6u ProSAT]</span></td></tr> | + | <tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[https://proteopedia.org/fgij/fg.htm?mol=6a6u FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=6a6u OCA], [https://pdbe.org/6a6u PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=6a6u RCSB], [https://www.ebi.ac.uk/pdbsum/6a6u PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=6a6u ProSAT]</span></td></tr> |
| | </table> | | </table> |
| | <div style="background-color:#fffaf0;"> | | <div style="background-color:#fffaf0;"> |
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| | __TOC__ | | __TOC__ |
| | </StructureSection> | | </StructureSection> |
| - | [[Category: A. nidulans]] | + | [[Category: Aspergillus nidulans]] |
| | [[Category: Large Structures]] | | [[Category: Large Structures]] |
| - | [[Category: Hashimoto, W]] | + | [[Category: Hashimoto W]] |
| - | [[Category: Itoh, T]] | + | [[Category: Itoh T]] |
| - | [[Category: Maruyama, Y]] | + | [[Category: Maruyama Y]] |
| - | [[Category: Murata, K]] | + | [[Category: Murata K]] |
| - | [[Category: Ogawa, N]] | + | [[Category: Ogawa N]] |
| - | [[Category: Aspergillus nidulan]]
| + | |
| - | [[Category: Fad-binding protein]]
| + | |
| - | [[Category: Fructosyl peptide]]
| + | |
| - | [[Category: Oxidoreductase]]
| + | |
| Structural highlights
6a6u is a 1 chain structure with sequence from Aspergillus nidulans. Full crystallographic information is available from OCA. For a guided tour on the structure components use FirstGlance.
| | Method: | X-ray diffraction, Resolution 1.945Å |
| Ligands: | , , , , |
| Resources: | FirstGlance, OCA, PDBe, RCSB, PDBsum, ProSAT |
Publication Abstract from PubMed
The currently available haemoglobin A1c (HbA1c) enzymatic assay consists of two specific steps: proteolysis of HbA1c and oxidation of the liberated fructosyl peptide by fructosyl peptide oxidase (FPOX). To develop a more convenient and high throughput assay, we devised novel protease-free assay system employing modified FPOX with HbA1c oxidation activity, namely HbA1c direct oxidase (HbA1cOX). AnFPOX-15, a modified FPOX from Aspergillus nidulans, was selected for conversion to HbA1cOX. As deduced from the crystal structure of AnFPOX-15, R61 was expected to obstruct the entrance of bulky substrates. An R61G mutant was thus constructed to open the gate at the active site. The prepared mutant exhibited significant reactivity for fructosyl hexapeptide (F-6P, N-terminal amino acids of HbA1c), and its crystal structure revealed a wider gate observed for AnFPOX-15. To improve the reactivity for F-6P, several mutagenesis approaches were performed. The ultimately generated AnFPOX-47 exhibited the highest F-6P reactivity and possessed HbA1c oxidation activity. HbA1c levels in blood samples as measured using the direct assay system using AnFPOX-47 were highly correlated with the levels measured using the conventional HPLC method. In this study, FPOX was successfully converted to HbA1cOX, which could represent a novel in vitro diagnostic modality for diabetes mellitus.
Creation of haemoglobin A1c direct oxidase from fructosyl peptide oxidase by combined structure-based site specific mutagenesis and random mutagenesis.,Ogawa N, Kimura T, Umehara F, Katayama Y, Nagai G, Suzuki K, Aisaka K, Maruyama Y, Itoh T, Hashimoto W, Murata K, Ichimura M Sci Rep. 2019 Jan 30;9(1):942. doi: 10.1038/s41598-018-37806-x. PMID:30700768[1]
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.
References
- ↑ Ogawa N, Kimura T, Umehara F, Katayama Y, Nagai G, Suzuki K, Aisaka K, Maruyama Y, Itoh T, Hashimoto W, Murata K, Ichimura M. Creation of haemoglobin A1c direct oxidase from fructosyl peptide oxidase by combined structure-based site specific mutagenesis and random mutagenesis. Sci Rep. 2019 Jan 30;9(1):942. doi: 10.1038/s41598-018-37806-x. PMID:30700768 doi:http://dx.doi.org/10.1038/s41598-018-37806-x
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