Modular structure of neuronal nitric oxide synthase: Localization of the arginine binding site and modulation by pterin

Jonathan S. Nishimura, Pavel Martasek, Kirk McMillan, John C. Salerno, Qing Liu, Steven S. Gross, Bettie Sue Siler Masters

Research output: Contribution to journalArticlepeer-review

50 Scopus citations

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

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

Fingerprint Dive into the research topics of 'Modular structure of neuronal nitric oxide synthase: Localization of the arginine binding site and modulation by pterin'. Together they form a unique fingerprint.

Cite this