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Related Experiment Videos

Nitric oxide scavenging by the cobalamin precursor cobinamide.

Kate E Broderick1, Veena Singh, Shunhui Zhuang

  • 1Department of Medicine and Cancer Center, University of California, San Diego, La Jolla, California 92093-0652, USA.

The Journal of Biological Chemistry
|January 6, 2005
PubMed
Summary

Cobinamide effectively scavenges nitric oxide (NO) both inside and outside cells. Its toxicity in biological systems can be eliminated by adding cobalamin (vitamin B12).

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Area of Science:

  • Biochemistry
  • Physiology
  • Pharmacology

Background:

  • Nitric oxide (NO) is a crucial signaling molecule.
  • NO synthesis inhibitors and scavengers are vital for studying NO functions and treating diseases with excess NO.
  • Cobinamide, a vitamin B12 precursor, has previously shown high affinity for NO.

Purpose of the Study:

  • To evaluate cobinamide's potential as a NO scavenger in biological systems.
  • To investigate cobinamide's efficacy in both in vitro and in vivo models.
  • To assess cobinamide's toxicity and the role of cobalamin in mitigating it.

Main Methods:

  • Testing cobinamide's effect on NO-stimulated fluid secretion in Drosophila Malpighian tubules.
  • Assessing cobinamide's impact on NO-induced VASP phosphorylation in cultured rodent and human cells.

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  • Measuring nitrite and nitrate concentrations to confirm NO scavenging.
  • Evaluating cobinamide's toxicity and its interaction with cobalamin in mammalian cells.
  • Investigating cobinamide's effect on cobalamin-dependent enzymes in vitro and in vivo.

    • Main Results:

    • Cobinamide reversed NO-stimulated fluid secretion in Drosophila.
    • Cobinamide prevented NO-induced VASP phosphorylation in cultured cells.
    • Cobinamide reduced nitrite and nitrate levels, confirming NO scavenging.
    • Cobinamide exhibited toxicity at concentrations >= 50 microM, which was prevented by cobalamin.
    • Cobinamide interfered with in vivo cobalamin-dependent enzyme activity, but not in the presence of cobalamin.

    Conclusions:

    • Cobinamide is an effective intra- and extracellular NO scavenger.
    • Cobinamide's toxicity is linked to interference with cobalamin metabolism and can be eliminated by co-administration of cobalamin.
    • A facile method for cobinamide production and purification was developed.