Quercetinase, a dioxygenase containing copper

Oka, Takuji; Simpson, F. J.

doi:10.1016/s0006-291x(71)80076-1

Cited by 103 publications

(55 citation statements)

References 11 publications

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“…Thus, whether pirin Sm functions as an enzyme, a regulator involved in any signaling pathway, or a protein enhancing the interaction between DNA and protein complexes remains to be further characterized among bacterial species. The S. marcescens pirin shows sequential and structural similarity to E. coli YhhW (20); however, whether pirin Sm is secreted outside the S. marcescens cells and degrades quercetin remains to be determined. In this report, through protein-protein interaction, genetic, and biochemical analyses, our results showed that pirin Sm binds with PDH E1 and inhibits PDH E1 and PDH enzyme complex activities.…”

Section: Discussionmentioning

confidence: 99%

Pirin Regulates Pyruvate Catabolism by Interacting with the Pyruvate Dehydrogenase E1 Subunit and Modulating Pyruvate Dehydrogenase Activity

et al. 2007

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The protein pirin, which is involved in a variety of biological processes, is conserved from prokaryotic microorganisms, fungi, and plants to mammals. It acts as a transcriptional cofactor or an apoptosis-related protein in mammals and is involved in seed germination and seedling development in plants. In prokaryotes, while pirin is stress induced in cyanobacteria and may act as a quercetinase in Escherichia coli, the functions of pirin orthologs remain mostly uncharacterized. We show that the Serratia marcescens pirin (pirin Sm ) gene encodes an ortholog of pirin protein. Protein pull-down and bacterial two-hybrid assays followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and electrospray ionization-tandem mass spectrometry analyses showed the pyruvate dehydrogenase (PDH) E1 subunit as a component interacting with the pirin Sm gene. Functional analyses showed that both PDH E1 subunit activity and PDH enzyme complex activity are inhibited by the pirin Sm gene in S. marcescens CH-1. The S. marcescens CH-1 pirin Sm gene was subsequently mutated by insertion-deletion homologous recombination. Accordingly, the PDH E1 and PDH enzyme complex activities and cellular ATP concentration increased up to 250%, 140%, and 220%, respectively, in the S. marcescens CH-1 pirin Sm mutant. Concomitantly, the cellular NADH/NAD ؉ ratio increased in the pirin Sm mutant, indicating increased tricarboxylic acid (TCA) cycle activity. Our results show that the pirin Sm gene plays a regulatory role in the process of pyruvate catabolism to acetyl coenzyme A through interaction with the PDH E1 subunit and inhibiting PDH enzyme complex activity in S. marcescens CH-1, and they suggest that pirin Sm is an important protein involved in determining the direction of pyruvate metabolism towards either the TCA cycle or the fermentation pathways.The protein pirin is widely found in mammals, plants, fungi, and also prokaryotic organisms (32). While the cellular functions of pirin show diversity and pirin homologs play important roles in a number of different biological processes, cellular localization of pirin is not restricted to specific compartments. In eukaryotes, pirin was initially isolated through a yeast twohybrid screen from the HeLa cell cDNA library and is localized within cell nuclei; it acts as an interactor with nuclear factor

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Section: Discussionmentioning

confidence: 99%

Pirin Regulates Pyruvate Catabolism by Interacting with the Pyruvate Dehydrogenase E1 Subunit and Modulating Pyruvate Dehydrogenase Activity

et al. 2007

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“…All quercetinases isolated from fungal sources (the Aspergillus and Penicillium enzymes) contain ~ 1 Cu atom per monomer, 1111,1112,1117 excepting A. niger 2,4-QD, which binds ~2 Cu per trimer. 1110 The copper is in the Cu II redox state, since a cuproine assay by Oka and Simpson detected very little Cu I in the as isolated enzyme.…”

Section: Substrate Activation By Cuii Sitesmentioning

confidence: 99%

“…1110 The copper is in the Cu II redox state, since a cuproine assay by Oka and Simpson detected very little Cu I in the as isolated enzyme. 1117 A. niger 2,4-QD is also isolated with variable Ni content in addition to Cu II , but the amount of nickel present does not affect the enzyme activity. 1110 In contrast, the prokaryote quercetinases from B. subtillis and Streptomyces are expressed in E. coli and bind many different divalent metals, depending on the metal content of the culture media.…”

Section: Substrate Activation By Cuii Sitesmentioning

confidence: 99%

Copper Active Sites in Biology

et al. 2014

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“…The latter compounds are known not only to chelate Fe(II) but also to form complexes with, e.g., Cu(II) (34,35). Therefore, ethylxanthate, highly specific for copper at low concentrations (36), was also applied. The latter clearly showed significant inhibition of the reaction at low concentrations of even 0.2 mM.…”

Section: Purification Of Lcp1mentioning

confidence: 99%

Latex Clearing Protein—an Oxygenase Cleaving Poly( cis -1,4-Isoprene) Rubber at the cis Double Bonds

Hiessl

Böse

Oetermann

et al. 2014

Appl Environ Microbiol

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dGordonia polyisoprenivorans strain VH2, a potent rubber-degrading actinomycete, harbors two latex clearing proteins (Lcps), which are known to be essential for the microbial degradation of rubber. However, biochemical information on the exact role of this protein in the degradation of polyisoprene was lacking. In this study, the gene encoding Lcp1 VH2 was heterologously expressed in strains of Escherichia coli, the corresponding protein was purified, and its role in rubber degradation was examined by measurement of oxygen consumption as well as by chromatographic and spectroscopic methods. It turned out that active Lcp1 VH2 is a monomer and is responsible for the oxidative cleavage of poly(cis-1,4-isoprene) in synthetic as well as in natural rubber by the addition of oxygen (O 2 ) to the cis double bonds. The resulting oligomers possess repetitive isoprene units with aldehyde (CHO-CH 2 -) and ketone (-CH 2 -CO-CH 3 ) functional groups at the termini. Two fractions with average isoprene contents of 18 and 10, respectively, were isolated, thus indicating an endocleavage mechanism. The activity of Lcp1 VH2 was determined by applying a polarographic assay. Alkenes, acyclic terpenes, or other rubber-like polymers, such as poly(cis-1,4-butadiene) or poly(trans-1,4-isoprene), are not oxidatively cleaved by Lcp1 VH2 . The pH and temperature optima of the enzyme are at pH 7 and 30°C, respectively. Furthermore, it was demonstrated that active Lcp1 VH2 is a Cu(II)-containing oxygenase that exhibits a conserved domain of unknown function which cannot be detected in any other hitherto-characterized enzyme. The results presented here indicate that this domain might represent a new protein family of oxygenases.

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Quercetinase, a dioxygenase containing copper

Cited by 103 publications

References 11 publications

Pirin Regulates Pyruvate Catabolism by Interacting with the Pyruvate Dehydrogenase E1 Subunit and Modulating Pyruvate Dehydrogenase Activity

Pirin Regulates Pyruvate Catabolism by Interacting with the Pyruvate Dehydrogenase E1 Subunit and Modulating Pyruvate Dehydrogenase Activity

Copper Active Sites in Biology

Latex Clearing Protein—an Oxygenase Cleaving Poly( cis -1,4-Isoprene) Rubber at the cis Double Bonds

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