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==Crystal structure of S-nitrosylated nitrosoglutathione reductase(GSNOR)from Chlamydomonas reinhardtii, in complex with NAD+== | ==Crystal structure of S-nitrosylated nitrosoglutathione reductase(GSNOR)from Chlamydomonas reinhardtii, in complex with NAD+== | ||
<StructureSection load='7av7' size='340' side='right'caption='[[7av7]]' scene=''> | <StructureSection load='7av7' size='340' side='right'caption='[[7av7]], [[Resolution|resolution]] 2.90Å' scene=''> | ||
== Structural highlights == | == Structural highlights == | ||
<table><tr><td colspan='2'>Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=7AV7 OCA]. For a <b>guided tour on the structure components</b> use [ | <table><tr><td colspan='2'>[[7av7]] is a 6 chain structure with sequence from [https://en.wikipedia.org/wiki/Chlamydomonas_reinhardtii Chlamydomonas reinhardtii]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=7AV7 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=7AV7 FirstGlance]. <br> | ||
</td></tr><tr id='resources'><td class="sblockLbl"><b>Resources:</b></td><td class="sblockDat"><span class='plainlinks'>[ | </td></tr><tr id='method'><td class="sblockLbl"><b>[[Empirical_models|Method:]]</b></td><td class="sblockDat" id="methodDat">X-ray diffraction, [[Resolution|Resolution]] 2.9Å</td></tr> | ||
<tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=CL:CHLORIDE+ION'>CL</scene>, <scene name='pdbligand=NAD:NICOTINAMIDE-ADENINE-DINUCLEOTIDE'>NAD</scene>, <scene name='pdbligand=SNC:S-NITROSO-CYSTEINE'>SNC</scene>, <scene name='pdbligand=ZN:ZINC+ION'>ZN</scene></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=7av7 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=7av7 OCA], [https://pdbe.org/7av7 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=7av7 RCSB], [https://www.ebi.ac.uk/pdbsum/7av7 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=7av7 ProSAT]</span></td></tr> | |||
</table> | </table> | ||
== Function == | |||
[https://www.uniprot.org/uniprot/A0A2K3D6R4_CHLRE A0A2K3D6R4_CHLRE] | |||
<div style="background-color:#fffaf0;"> | |||
== Publication Abstract from PubMed == | |||
Protein S-nitrosylation plays a fundamental role in cell signaling and nitrosoglutathione (GSNO) is considered as the main nitrosylating signaling molecule. Enzymatic systems controlling GSNO homeostasis are thus crucial to indirectly control the formation of protein S-nitrosothiols. GSNO reductase (GSNOR) is the key enzyme controlling GSNO levels by catalyzing its degradation in the presence of NADH. Here, we found that protein extracts from the microalga Chlamydomonas reinhardtii catabolize GSNO via two enzymatic systems having specific reliance on NADPH or NADH and different biochemical features. Scoring the Chlamydomonas genome for orthologs of known plant GSNORs, we found two genes encoding for putative and almost identical GSNOR isoenzymes. One of the two, here named CrGSNOR1, was heterologously expressed and purified. Its kinetic properties were determined and the three-dimensional structures of the apo-, NAD(+)- and NAD(+)/GSNO-forms were solved. These analyses revealed that CrGSNOR1 has a strict specificity towards GSNO and NADH, and a conserved folding with respect to other plant GSNORs. The catalytic zinc ion, however, showed an unexpected variability of the coordination environment. Furthermore, we evaluated the catalytic response of CrGSNOR1 to thermal denaturation, thiol-modifying agents and oxidative modifications as well as the reactivity and position of accessible cysteines. Despite being a cysteine-rich protein, CrGSNOR1 contains only two solvent-exposed/reactive cysteines. Oxidizing and nitrosylating treatments have null or limited effects on CrGSNOR1 activity and folding, highlighting a certain resistance of the algal enzyme to redox modifications. The molecular mechanisms and structural features underlying the response to thiol-based modifications are discussed. | |||
Structural and functional insights into nitrosoglutathione reductase from Chlamydomonas reinhardtii.,Tagliani A, Rossi J, Marchand CH, De Mia M, Tedesco D, Gurrieri L, Meloni M, Falini G, Trost P, Lemaire SD, Fermani S, Zaffagnini M Redox Biol. 2020 Nov 24;38:101806. doi: 10.1016/j.redox.2020.101806. PMID:33316743<ref>PMID:33316743</ref> | |||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |||
</div> | |||
<div class="pdbe-citations 7av7" style="background-color:#fffaf0;"></div> | |||
== References == | |||
<references/> | |||
__TOC__ | __TOC__ | ||
</StructureSection> | </StructureSection> | ||
[[Category: Chlamydomonas reinhardtii]] | |||
[[Category: Large Structures]] | [[Category: Large Structures]] | ||
[[Category: Falini G]] | [[Category: Falini G]] | ||
Latest revision as of 15:16, 1 February 2024
Crystal structure of S-nitrosylated nitrosoglutathione reductase(GSNOR)from Chlamydomonas reinhardtii, in complex with NAD+Crystal structure of S-nitrosylated nitrosoglutathione reductase(GSNOR)from Chlamydomonas reinhardtii, in complex with NAD+
Structural highlights
FunctionPublication Abstract from PubMedProtein S-nitrosylation plays a fundamental role in cell signaling and nitrosoglutathione (GSNO) is considered as the main nitrosylating signaling molecule. Enzymatic systems controlling GSNO homeostasis are thus crucial to indirectly control the formation of protein S-nitrosothiols. GSNO reductase (GSNOR) is the key enzyme controlling GSNO levels by catalyzing its degradation in the presence of NADH. Here, we found that protein extracts from the microalga Chlamydomonas reinhardtii catabolize GSNO via two enzymatic systems having specific reliance on NADPH or NADH and different biochemical features. Scoring the Chlamydomonas genome for orthologs of known plant GSNORs, we found two genes encoding for putative and almost identical GSNOR isoenzymes. One of the two, here named CrGSNOR1, was heterologously expressed and purified. Its kinetic properties were determined and the three-dimensional structures of the apo-, NAD(+)- and NAD(+)/GSNO-forms were solved. These analyses revealed that CrGSNOR1 has a strict specificity towards GSNO and NADH, and a conserved folding with respect to other plant GSNORs. The catalytic zinc ion, however, showed an unexpected variability of the coordination environment. Furthermore, we evaluated the catalytic response of CrGSNOR1 to thermal denaturation, thiol-modifying agents and oxidative modifications as well as the reactivity and position of accessible cysteines. Despite being a cysteine-rich protein, CrGSNOR1 contains only two solvent-exposed/reactive cysteines. Oxidizing and nitrosylating treatments have null or limited effects on CrGSNOR1 activity and folding, highlighting a certain resistance of the algal enzyme to redox modifications. The molecular mechanisms and structural features underlying the response to thiol-based modifications are discussed. Structural and functional insights into nitrosoglutathione reductase from Chlamydomonas reinhardtii.,Tagliani A, Rossi J, Marchand CH, De Mia M, Tedesco D, Gurrieri L, Meloni M, Falini G, Trost P, Lemaire SD, Fermani S, Zaffagnini M Redox Biol. 2020 Nov 24;38:101806. doi: 10.1016/j.redox.2020.101806. PMID:33316743[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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