Development of a dimethylarginine dimethylaminohydrolase (DDAH) assay for high-throughput chemical screening

Yohannes T. Ghebremariam, Daniel A. Erlanson, Keisuke Yamada, John P. Cooke

Research output: Contribution to journalArticle

16 Scopus citations

Abstract

Nitric oxide (NO) is a potent signaling molecule that needs to be tightly regulated to maintain metabolic and cardiovascular homeostasis. The nitric oxide synthase (NOS)/dimethylarginine dimethylaminohydrolase (DDAH)/asymmetric dimethylarginine (ADMA) pathway is central to this regulation. Specifically, the small-molecule ADMA competitively inhibits NOS, thus lowering NO levels. The majority of ADMA is physiologically metabolized by DDAH, thus maintaining NO levels at a physiological concentration. However, under pathophysiological conditions, DDAH activity is impaired, in part as a result of its sensitivity to oxidative stress. Therefore, the application of high-throughput chemical screening for the discovery of small molecules that could restore or enhance DDAH activity might have significant potential in treating metabolic and vascular diseases characterized by reduced NO levels, including atherosclerosis, hypertension, and insulin resistance. By contrast, excessive generation of NO (primarily driven by inducible NOS) could play a role in idiopathic pulmonary fibrosis, sepsis, migraine headaches, and some types of cancer. In these conditions, small molecules that inhibit DDAH activity might be therapeutically useful. Here, we describe optimization and validation of a highly reproducible and robust assay successfully used in a high-throughput screen for DDAH modulators.

Original languageEnglish (US)
Pages (from-to)651-661
Number of pages11
JournalJournal of Biomolecular Screening
Volume17
Issue number5
DOIs
StatePublished - Jun 2012

Keywords

  • asymmetric dimethylarginine
  • diabetes
  • hypertension
  • idiopathic pulmonary fibrosis
  • nitric oxide

ASJC Scopus subject areas

  • Analytical Chemistry
  • Drug Discovery
  • Pharmacology
  • Biochemistry
  • Molecular Medicine
  • Biotechnology

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