7kw9: Difference between revisions
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<StructureSection load='7kw9' size='340' side='right'caption='[[7kw9]]' scene=''> | <StructureSection load='7kw9' size='340' side='right'caption='[[7kw9]]' scene=''> | ||
== Structural highlights == | == Structural highlights == | ||
<table><tr><td colspan='2'>Full | <table><tr><td colspan='2'>[[7kw9]] is a 1 chain structure with sequence from [https://en.wikipedia.org/wiki/Runella_slithyformis Runella slithyformis]. Full experimental information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=7KW9 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=7KW9 FirstGlance]. <br> | ||
</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=7kw9 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=7kw9 OCA], [https://pdbe.org/7kw9 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=7kw9 RCSB], [https://www.ebi.ac.uk/pdbsum/7kw9 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=7kw9 ProSAT]</span></td></tr> | </td></tr><tr id='ligand'><td class="sblockLbl"><b>[[Ligand|Ligands:]]</b></td><td class="sblockDat" id="ligandDat"><scene name='pdbligand=NAD:NICOTINAMIDE-ADENINE-DINUCLEOTIDE'>NAD</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=7kw9 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=7kw9 OCA], [https://pdbe.org/7kw9 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=7kw9 RCSB], [https://www.ebi.ac.uk/pdbsum/7kw9 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=7kw9 ProSAT]</span></td></tr> | |||
</table> | </table> | ||
== Function == | |||
[https://www.uniprot.org/uniprot/A0A7U3ZK32_RUNSL A0A7U3ZK32_RUNSL] Removes the 2'-phosphate from RNA via an intermediate in which the phosphate is ADP-ribosylated by NAD followed by a presumed transesterification to release the RNA and generate ADP-ribose 1''-2''-cyclic phosphate (APPR>P). May function as an ADP-ribosylase.[ARBA:ARBA00025212][HAMAP-Rule:MF_00299] | |||
<div style="background-color:#fffaf0;"> | |||
== Publication Abstract from PubMed == | |||
Tpt1, an essential component of the fungal and plant tRNA splicing machinery, catalyzes transfer of an internal RNA 2'-PO4 to NAD+ yielding RNA 2'-OH and ADP-ribose-1',2'-cyclic phosphate products. Here, we report NMR structures of the Tpt1 ortholog from the bacterium Runella slithyformis (RslTpt1), as apoenzyme and bound to NAD+. RslTpt1 consists of N- and C-terminal lobes with substantial inter-lobe dynamics in the free and NAD+-bound states. ITC measurements of RslTpt1 binding to NAD+ (KD approximately 31 muM), ADP-ribose ( approximately 96 muM) and ADP ( approximately 123 muM) indicate that substrate affinity is determined primarily by the ADP moiety; no binding of NMN or nicotinamide is observed by ITC. NAD+-induced chemical shift perturbations (CSPs) localize exclusively to the RslTpt1 C-lobe. NADP+, which contains an adenylate 2'-PO4 (mimicking the substrate RNA 2'-PO4), binds with lower affinity (KD approximately 1 mM) and elicits only N-lobe CSPs. The RslTpt1.NAD+ binary complex reveals C-lobe contacts to adenosine ribose hydroxyls (His99, Thr101), the adenine nucleobase (Asn105, Asp112, Gly113, Met117) and the nicotinamide riboside (Ser125, Gln126, Asn163, Val165), several of which are essential for RslTpt1 activity in vivo. Proximity of the NAD+ beta-phosphate to ribose-C1'' suggests that it may stabilize an oxocarbenium transition-state during the first step of the Tpt1-catalyzed reaction. | |||
NMR solution structures of Runella slithyformis RNA 2'-phosphotransferase Tpt1 provide insights into NAD+ binding and specificity.,Alphonse S, Banerjee A, Dantuluri S, Shuman S, Ghose R Nucleic Acids Res. 2021 Sep 27;49(17):9607-9624. doi: 10.1093/nar/gkab241. PMID:33880546<ref>PMID:33880546</ref> | |||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |||
</div> | |||
<div class="pdbe-citations 7kw9" style="background-color:#fffaf0;"></div> | |||
== References == | |||
<references/> | |||
__TOC__ | __TOC__ | ||
</StructureSection> | </StructureSection> | ||
[[Category: Large Structures]] | [[Category: Large Structures]] | ||
[[Category: Runella slithyformis]] | |||
[[Category: Alphonse S]] | [[Category: Alphonse S]] | ||
[[Category: Banerjee A]] | [[Category: Banerjee A]] | ||
Latest revision as of 14:14, 14 June 2023
NMR Structure of a tRNA 2'-phosphotransferase from Runella slithyformis in complex with NAD+NMR Structure of a tRNA 2'-phosphotransferase from Runella slithyformis in complex with NAD+
Structural highlights
FunctionA0A7U3ZK32_RUNSL Removes the 2'-phosphate from RNA via an intermediate in which the phosphate is ADP-ribosylated by NAD followed by a presumed transesterification to release the RNA and generate ADP-ribose 1-2-cyclic phosphate (APPR>P). May function as an ADP-ribosylase.[ARBA:ARBA00025212][HAMAP-Rule:MF_00299] Publication Abstract from PubMedTpt1, an essential component of the fungal and plant tRNA splicing machinery, catalyzes transfer of an internal RNA 2'-PO4 to NAD+ yielding RNA 2'-OH and ADP-ribose-1',2'-cyclic phosphate products. Here, we report NMR structures of the Tpt1 ortholog from the bacterium Runella slithyformis (RslTpt1), as apoenzyme and bound to NAD+. RslTpt1 consists of N- and C-terminal lobes with substantial inter-lobe dynamics in the free and NAD+-bound states. ITC measurements of RslTpt1 binding to NAD+ (KD approximately 31 muM), ADP-ribose ( approximately 96 muM) and ADP ( approximately 123 muM) indicate that substrate affinity is determined primarily by the ADP moiety; no binding of NMN or nicotinamide is observed by ITC. NAD+-induced chemical shift perturbations (CSPs) localize exclusively to the RslTpt1 C-lobe. NADP+, which contains an adenylate 2'-PO4 (mimicking the substrate RNA 2'-PO4), binds with lower affinity (KD approximately 1 mM) and elicits only N-lobe CSPs. The RslTpt1.NAD+ binary complex reveals C-lobe contacts to adenosine ribose hydroxyls (His99, Thr101), the adenine nucleobase (Asn105, Asp112, Gly113, Met117) and the nicotinamide riboside (Ser125, Gln126, Asn163, Val165), several of which are essential for RslTpt1 activity in vivo. Proximity of the NAD+ beta-phosphate to ribose-C1 suggests that it may stabilize an oxocarbenium transition-state during the first step of the Tpt1-catalyzed reaction. NMR solution structures of Runella slithyformis RNA 2'-phosphotransferase Tpt1 provide insights into NAD+ binding and specificity.,Alphonse S, Banerjee A, Dantuluri S, Shuman S, Ghose R Nucleic Acids Res. 2021 Sep 27;49(17):9607-9624. doi: 10.1093/nar/gkab241. PMID:33880546[1] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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