Use of chimeric forms of neuronal nitric-oxide synthase as dominant negative mutants

Yume T. Phung, Stephen Matthew Black

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Because the functional form of neuronal nitric-oxide synthase (nNOS) is a homodimer, we investigated whether we could disrupt dimer formation with inactive nNOS chimeras acting as dominant negative mutants. To test this hypothesis, we either expressed the heme and reductase regions of rat nNOS as single domains or produced fusion proteins between the rat nNOS heme domain and various other electron-shuttling proteins. A dominant negative potential of these constructs was demonstrated by their ability to reduce NOS activity when transfected into a cell line stably expressing rat nNOS. In the presence of these nNOS mutant proteins, cellular levels of inactive nNOS monomers were significantly increased, indicating that their mechanism of action is through the disruption of nNOS dimer formation. These dominant negative mutants should prove valuable in analyzing the role of nNOS in biological systems.

Original languageEnglish (US)
Pages (from-to)333-338
Number of pages6
JournalIUBMB Life
Volume48
Issue number3
DOIs
StatePublished - Nov 17 1999

Fingerprint

Nitric Oxide Synthase Type I
Rats
Heme
Dimers
Biological systems
Mutant Proteins
Oxidoreductases
Proteins
Fusion reactions
Monomers
Cells
Electrons
Cell Line

Keywords

  • Dimer
  • Dominant negative
  • Expression
  • Nitric oxide

ASJC Scopus subject areas

  • Biochemistry
  • Molecular Biology
  • Genetics
  • Clinical Biochemistry
  • Cell Biology

Cite this

Use of chimeric forms of neuronal nitric-oxide synthase as dominant negative mutants. / Phung, Yume T.; Black, Stephen Matthew.

In: IUBMB Life, Vol. 48, No. 3, 17.11.1999, p. 333-338.

Research output: Contribution to journalArticle

Phung, Yume T. ; Black, Stephen Matthew. / Use of chimeric forms of neuronal nitric-oxide synthase as dominant negative mutants. In: IUBMB Life. 1999 ; Vol. 48, No. 3. pp. 333-338.
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