Computational and experimental studies on the catalytic mechanism of biliverdin-IXbeta reductase

Liam J Smith, Seamus Browne, Adrian J Mulholland, Timothy J Mantle

Research output: Contribution to journalArticle (Academic Journal)peer-review

31 Citations (Scopus)


BVR-B (biliverdin-IXbeta reductase) also known as FR (flavin reductase) is a promiscuous enzyme catalysing the pyridine-nucleotide-dependent reduction of a variety of flavins, biliverdins, PQQ (pyrroloquinoline quinone) and ferric ion. Mechanistically it is a good model for BVR-A (biliverdin-IXalpha reductase), a potential pharmacological target for neonatal jaundice and also a potential target for adjunct therapy to maintain protective levels of biliverdin-IXalpha during organ transplantation. In a commentary on the structure of BVR-B it was noted that one outstanding issue remained: whether the mechanism was a concerted hydride transfer followed by protonation of a pyrrolic anion or protonation of the pyrrole followed by hydride transfer. In the present study we have attempted to address this question using QM/MM (quantum mechanics/molecular mechanics) calculations. QM/MM potential energy surfaces show that the lowest energy pathway proceeds with a positively charged pyrrole intermediate via two transition states. These initial calculations were performed with His(153) as the source of the proton. However site-directed mutagenesis studies with both the H153A and the H153N mutant reveal that His(153) is not required for catalytic activity. We have repeated the calculation with a solvent hydroxonium donor and obtain a similar energy landscape indicating that protonation of the pyrrole is the most likely first step followed by hydride transfer and that the required proton may come from bulk solvent. The implications of the present study for the design of inhibitors of BVR-A are discussed.

Original languageEnglish
Pages (from-to)475-84
Number of pages10
JournalBiochemical Journal
Issue number3
Publication statusPublished - 1 May 2008


  • Biliverdine
  • Catalysis
  • Computer Simulation
  • Crystallography, X-Ray
  • Humans
  • Kinetics
  • Models, Molecular
  • Molecular Structure
  • Mutation
  • NADP
  • Oxidoreductases Acting on CH-CH Group Donors
  • Protein Binding
  • Protons
  • Substrate Specificity


Dive into the research topics of 'Computational and experimental studies on the catalytic mechanism of biliverdin-IXbeta reductase'. Together they form a unique fingerprint.

Cite this