6RG0

Structure of pdxj

Summary for 6RG0

• Entry DOI: 10.2210/pdb6rg0/pdb
• Descriptor: Pyridoxine 5'-phosphate synthase (1 entity in total)
• Functional Keywords: serine phosphatase serb, serine aminotransferase serc, histidinol phosphatase hisb, histidinol phosphatase aminotransferase hisc, transferase
• Biological source: Escherichia coli K12
• Total number of polymer chains: 4
• Total formula weight: 116906.72

Authors

Rohweder, B.,Rajendran, C.,Sterner, R. (deposition date: 2019-04-16, release date: 2020-05-13, Last modification date: 2024-01-24)

Primary citation

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.
Library Selection with a Randomized Repertoire of ( beta alpha )8-Barrel Enzymes Results in Unexpected Induction of Gene Expression.
Biochemistry, 58:4207-4217, 2019

PubMed Abstract: The potential of the frequently encountered (βα)-barrel fold to acquire new functions was tested by an approach combining random mutagenesis and selection . For this purpose, the genes encoding 52 different phosphate-binding (βα)-barrel proteins were subjected to error-prone PCR and cloned into an expression plasmid. The resulting mixed repertoire was used to transform different auxotrophic 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 genes E (encoding a putative -acetylmannosamine-6-phosphate 2-epimerase) and J (encoding the pyridoxine 5'-phosphate synthase) were responsible for rescuing both ΔB and ΔC. Unexpectedly, the complementing NanE and PdxJ variants did not catalyze the SerB or SerC reactions . Instead, RT-qPCR, RNAseq, and transcriptome analysis showed that they rescue the deletions by enlisting the help of endogenous 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 ΔB and ΔC through point mutations and recruitment of additional amino acids in NanE and PdxJ provides another example for the adaptability of the (βα)-barrel fold.

PubMed: 31557000
DOI: 10.1021/acs.biochem.9b00579

Experimental method

X-RAY DIFFRACTION (3.074 Å)