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Studies on the incorporation of labelled sulphate into cells and cell-free extracts of Nitrosomonas europaea

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Summary

The incorporation of [35S]sulphate was followed into the washed cell suspensions of Nitrosomonas europaea. Thus bound sulphate, sulphite, sulphide, cysteine, glutathione, homocysteine and methionine were found in the ethanol soluble fraction as well as in the residual hydrolysed protein fraction. Cysteic acid, methionine sulphoxide and methionine sulphone were detected in the residual protein. The reaction between sulphydryl groups and N-ethylmaleimide has been successfully used to stabilize the thiol compounds in cell-extracts and the derivatives thus obtained were separated by paper chromatography. As in other microorganisms, sulphate is first activated by ATP in Nitrosomonas before it is reduced. The formation of APS and PAPS has been studied. A pathway for the incorporation of [35S]sulphate is proposed.

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Abbreviations

POPOP:

1,4-bis-(5-phenyloxazolyl-2)-benzene

PPO:

2,5-diphenyloxazole

APS:

adenosine-5′-phosphosulphate

PAPS:

adenosine-3′-phosphate 5′-phosphosulphate

ATP:

adenosine triphosphate

DNA-ase:

deoxyribonuclease

NEM:

N-ethylmaleimide

TCA:

trichloro-acetic acid

GSH:

glutathione

References

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Varma, A.K., Nicholas, D.J.D. Studies on the incorporation of labelled sulphate into cells and cell-free extracts of Nitrosomonas europaea . Archiv. Mikrobiol. 73, 293–307 (1970). https://doi.org/10.1007/BF00412296

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Keywords

  • Methionine
  • Homocysteine
  • Hydrolyse Protein
  • Sulphone
  • Sulphite