Isolation and immunocytochemical location of the nitrite-oxidizing system in Nitrospira moscoviensis

E. Spieck, Silke Ehrich, Jens Aamand, Eberhard Bock

Publikation: Bidrag til tidsskriftArtikelForskningpeer review

61 Citationer (Scopus)

Abstrakt

A membrane-associated nitrite-oxidizing system of Nitrospira moscoviensis was isolated from heat-treated membranes. The four major proteins of the enzyme fraction had apparent molecular masses of 130, 62, 46, and 29 kDa, respectively. The nitrite-oxidizing activity was dependent on the presence of molybdenum. In contrast to the nitrite oxidoreductase of Nitrobacter hamburgensis X14, the activity of the nitrite-oxidizing system of Ns. moscoviensis increased when solubilized by heat treatment. Electron microscopy of the purified enzyme revealed uniform particles with a size of approximately 7 x 9 nm. SDS-immunoblotting analysis of crude extracts showed that the monoclonal antibodies Hyb 153-3, which recognize the β-subunit of the nitrite oxidoreductase from Nitrobacter, reacted with a protein of 50 kDa in Ns. moscoviensis. This protein corresponded to the protein of 46 kDa of the purified enzyme and contained a b-type cytochrome. Using electron microscopic immunocytochemistry and the monoclonal antibodies Hyb 153-3, the nitrite-oxidizing system of Ns. moscoviensis was shown to be located in the periplasmic space. Here a periodic arrangement of membrane-associated particles was found on the outside of the cytoplasmic membrane in the form of a hexagonal pattern. It is supposed that these particles represent the nitrite-oxidizing system in Nitrospira.

OriginalsprogEngelsk
Sider (fra-til)225-230
Antal sider6
TidsskriftArchives of Microbiology
Vol/bind169
Udgave nummer3
DOI
StatusUdgivet - feb. 1998

Emneord

  • Immunoassay
  • Ammonium
  • Oxidation

Programområde

  • Programområde 2: Vandressourcer

Fingeraftryk

Dyk ned i forskningsemnerne om 'Isolation and immunocytochemical location of the nitrite-oxidizing system in Nitrospira moscoviensis'. Sammen danner de et unikt fingeraftryk.

Citationsformater