Modular Structure of Neuronal Nitric Oxide Synthase: Localization of the Arginine Binding Site and Modulation by Pterin

J. S. Nishimura, P. Martasek, K. Mcmillan, J. C. Salerno, Q. Liu, S. S. Gross, B. S S Masters

Research output: Contribution to journalArticle

50 Citations (Scopus)

Abstract

A putative dihydrofolate reductase (DHFR) module has been identified in neuronal nitric oxide synthase, consisting of amino acids 558-721, and is proposed to be the site of tetrahydrobiopterin (BH4) binding. This polypeptide has been expressed in E. coli as a fusion protein with glutathione S-transferase (GST), using the plasmid pGEX-4T1. The protein binds Nω-nitro-L-arginine (NNA) tightly, but this binding is not stimulated by BH4. cDNAs for Module II (residues 220-557) and Module III (residues 220-721) have been expressed as fusion proteins with GST. Module II does not bind NNA. However, Module III does bind NNA and binding is significantly stimulated by BH4. These observations are taken as strong evidence that the DHFR module contains the L-arginine binding site and, presumably, the BH4 binding site by analogy to its homology with DHFR, but that tight binding of BH4 requires amino acids 220-577.

Original languageEnglish (US)
Pages (from-to)288-294
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume210
Issue number2
DOIs
StatePublished - May 16 1995

Fingerprint

Pterins
Nitric Oxide Synthase Type I
Tetrahydrofolate Dehydrogenase
Arginine
Binding Sites
Modulation
Glutathione Transferase
Fusion reactions
Amino Acids
Proteins
Escherichia coli
Plasmids
Complementary DNA
Peptides

ASJC Scopus subject areas

  • Molecular Biology
  • Cell Biology
  • Biophysics
  • Biochemistry

Cite this

Modular Structure of Neuronal Nitric Oxide Synthase : Localization of the Arginine Binding Site and Modulation by Pterin. / Nishimura, J. S.; Martasek, P.; Mcmillan, K.; Salerno, J. C.; Liu, Q.; Gross, S. S.; Masters, B. S S.

In: Biochemical and Biophysical Research Communications, Vol. 210, No. 2, 16.05.1995, p. 288-294.

Research output: Contribution to journalArticle

Nishimura, J. S. ; Martasek, P. ; Mcmillan, K. ; Salerno, J. C. ; Liu, Q. ; Gross, S. S. ; Masters, B. S S. / Modular Structure of Neuronal Nitric Oxide Synthase : Localization of the Arginine Binding Site and Modulation by Pterin. In: Biochemical and Biophysical Research Communications. 1995 ; Vol. 210, No. 2. pp. 288-294.
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