6RL0

Recombinant Pseudomonas stutzeri nitrous oxide reductase, form I

Summary for 6RL0

• Entry DOI: 10.2210/pdb6rl0/pdb
• Descriptor: Nitrous-oxide reductase, 2-[3-(2-HYDROXY-1,1-DIHYDROXYMETHYL-ETHYLAMINO)-PROPYLAMINO]-2-HYDROXYMETHYL-PROPANE-1,3-DIOL, CHLORIDE ION, ... (10 entities in total)
• Functional Keywords: denitrification, cofactor biogenesis, metal binding protein
• Biological source: Pseudomonas stutzeri
• Total number of polymer chains: 4
• Total formula weight: 287357.18

Authors

Zhang, L.,Wuest, A.,Prasser, B.,Mueller, C.,Einsle, O. (deposition date: 2019-04-30, release date: 2019-06-26, Last modification date: 2024-01-24)

Primary citation

Zhang, L.,Wust, A.,Prasser, B.,Muller, C.,Einsle, O.
Functional assembly of nitrous oxide reductase provides insights into copper site maturation.
Proc.Natl.Acad.Sci.USA, 116:12822-12827, 2019

PubMed Abstract: The multicopper enzyme nitrous oxide reductase reduces the greenhouse gas NO to uncritical N as the final step of bacterial denitrification. Its two metal centers require an elaborate assembly machinery that so far has precluded heterologous production as a prerequisite for bioremediatory applications in agriculture and wastewater treatment. Here, we report on the production of active holoenzyme in using a two-plasmid system to produce the entire biosynthetic machinery as well as the structural gene for the enzyme. Using this recombinant system to probe the role of individual maturation factors, we find that the ABC transporter NosFY and the accessory NosD protein are essential for the formation of the [4Cu:2S] site Cu, but not the electron transfer site Cu Depending on source organism, the heterologous host can, in some cases, compensate for the lack of the Cu chaperone NosL, while in others this protein is strictly required, underlining the case for designing a recombinant system to be entirely self-contained.

PubMed: 31189605
DOI: 10.1073/pnas.1903819116

Experimental method

X-RAY DIFFRACTION (1.78 Å)