6rg0: Difference between revisions
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==Structure of pdxj== | ==Structure of pdxj== | ||
<StructureSection load='6rg0' size='340' side='right'caption='[[6rg0]]' scene=''> | <StructureSection load='6rg0' size='340' side='right'caption='[[6rg0]], [[Resolution|resolution]] 3.07Å' 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=6RG0 OCA]. For a <b>guided tour on the structure components</b> use [ | <table><tr><td colspan='2'>[[6rg0]] is a 4 chain structure with sequence from [https://en.wikipedia.org/wiki/Escherichia_coli_K-12 Escherichia coli K-12]. Full crystallographic information is available from [http://oca.weizmann.ac.il/oca-bin/ocashort?id=6RG0 OCA]. For a <b>guided tour on the structure components</b> use [https://proteopedia.org/fgij/fg.htm?mol=6RG0 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]] 3.074Å</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=6rg0 FirstGlance], [http://oca.weizmann.ac.il/oca-bin/ocaids?id=6rg0 OCA], [https://pdbe.org/6rg0 PDBe], [https://www.rcsb.org/pdb/explore.do?structureId=6rg0 RCSB], [https://www.ebi.ac.uk/pdbsum/6rg0 PDBsum], [https://prosat.h-its.org/prosat/prosatexe?pdbcode=6rg0 ProSAT]</span></td></tr> | |||
</table> | </table> | ||
== Function == | |||
[https://www.uniprot.org/uniprot/PDXJ_ECOLI PDXJ_ECOLI] Catalyzes the complicated ring closure reaction between the two acyclic compounds 1-deoxy-D-xylulose-5-phosphate (DXP) and 3-amino-2-oxopropyl phosphate (1-amino-acetone-3-phosphate or AAP) to form pyridoxine 5'-phosphate (PNP) and inorganic phosphate.<ref>PMID:10225425</ref> | |||
<div style="background-color:#fffaf0;"> | |||
== Publication Abstract from PubMed == | |||
The potential of the frequently encountered (betaalpha)8-barrel fold to acquire new functions was tested by an approach combining random mutagenesis and selection in vivo. For this purpose, the genes encoding 52 different phosphate-binding (betaalpha)8-barrel proteins were subjected to error-prone PCR and cloned into an expression plasmid. The resulting mixed repertoire was used to transform different auxotrophic Escherichia coli strains, each lacking an enzyme with a phosphate-containing substrate. After plating of the different transformants on minimal medium, growth was observed only for two strains, lacking either the gene for the serine phosphatase SerB or the phosphoserine aminotransferase SerC. The same mutants of the E. coli genes nanE (encoding a putative N-acetylmannosamine-6-phosphate 2-epimerase) and pdxJ (encoding the pyridoxine 5'-phosphate synthase) were responsible for rescuing both DeltaserB and DeltaserC. Unexpectedly, the complementing NanE and PdxJ variants did not catalyze the SerB or SerC reactions in vitro. Instead, RT-qPCR, RNAseq, and transcriptome analysis showed that they rescue the deletions by enlisting the help of endogenous E. coli enzymes HisB and HisC through exclusive up-regulation of histidine operon transcription. While the promiscuous SerB activity of HisB is well-established, our data indicate that HisC is promiscuous for the SerC reaction, as well. The successful rescue of DeltaserB and DeltaserC through point mutations and recruitment of additional amino acids in NanE and PdxJ provides another example for the adaptability of the (betaalpha)8-barrel fold. | |||
Library Selection with a Randomized Repertoire of (betaalpha)8-Barrel Enzymes Results in Unexpected Induction of Gene Expression.,Rohweder B, Lehmann G, Eichner N, Polen T, Rajendran C, Ruperti F, Linde M, Treiber T, Jung O, Dettmer K, Meister G, Bott M, Gronwald W, Sterner R Biochemistry. 2019 Oct 15;58(41):4207-4217. doi: 10.1021/acs.biochem.9b00579., Epub 2019 Oct 7. PMID:31557000<ref>PMID:31557000</ref> | |||
From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine.<br> | |||
</div> | |||
<div class="pdbe-citations 6rg0" style="background-color:#fffaf0;"></div> | |||
== References == | |||
<references/> | |||
__TOC__ | __TOC__ | ||
</StructureSection> | </StructureSection> | ||
[[Category: Escherichia coli K-12]] | |||
[[Category: Large Structures]] | [[Category: Large Structures]] | ||
[[Category: Rajendran C]] | [[Category: Rajendran C]] | ||
[[Category: Rohweder B]] | [[Category: Rohweder B]] | ||
[[Category: Sterner R]] | [[Category: Sterner R]] | ||
Latest revision as of 15:19, 24 January 2024
Structure of pdxjStructure of pdxj
Structural highlights
FunctionPDXJ_ECOLI Catalyzes the complicated ring closure reaction between the two acyclic compounds 1-deoxy-D-xylulose-5-phosphate (DXP) and 3-amino-2-oxopropyl phosphate (1-amino-acetone-3-phosphate or AAP) to form pyridoxine 5'-phosphate (PNP) and inorganic phosphate.[1] Publication Abstract from PubMedThe potential of the frequently encountered (betaalpha)8-barrel fold to acquire new functions was tested by an approach combining random mutagenesis and selection in vivo. For this purpose, the genes encoding 52 different phosphate-binding (betaalpha)8-barrel proteins were subjected to error-prone PCR and cloned into an expression plasmid. The resulting mixed repertoire was used to transform different auxotrophic Escherichia coli strains, each lacking an enzyme with a phosphate-containing substrate. After plating of the different transformants on minimal medium, growth was observed only for two strains, lacking either the gene for the serine phosphatase SerB or the phosphoserine aminotransferase SerC. The same mutants of the E. coli genes nanE (encoding a putative N-acetylmannosamine-6-phosphate 2-epimerase) and pdxJ (encoding the pyridoxine 5'-phosphate synthase) were responsible for rescuing both DeltaserB and DeltaserC. Unexpectedly, the complementing NanE and PdxJ variants did not catalyze the SerB or SerC reactions in vitro. Instead, RT-qPCR, RNAseq, and transcriptome analysis showed that they rescue the deletions by enlisting the help of endogenous E. coli enzymes HisB and HisC through exclusive up-regulation of histidine operon transcription. While the promiscuous SerB activity of HisB is well-established, our data indicate that HisC is promiscuous for the SerC reaction, as well. The successful rescue of DeltaserB and DeltaserC through point mutations and recruitment of additional amino acids in NanE and PdxJ provides another example for the adaptability of the (betaalpha)8-barrel fold. Library Selection with a Randomized Repertoire of (betaalpha)8-Barrel Enzymes Results in Unexpected Induction of Gene Expression.,Rohweder B, Lehmann G, Eichner N, Polen T, Rajendran C, Ruperti F, Linde M, Treiber T, Jung O, Dettmer K, Meister G, Bott M, Gronwald W, Sterner R Biochemistry. 2019 Oct 15;58(41):4207-4217. doi: 10.1021/acs.biochem.9b00579., Epub 2019 Oct 7. PMID:31557000[2] From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine. References
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