Structure, dynamics and function of the evolutionarily changing biliverdin reductase B family

J Biochem. 2020 Aug 1;168(2):191-202. doi: 10.1093/jb/mvaa039.

Abstract

Biliverdin reductase B (BLVRB) family members are general flavin reductases critical in maintaining cellular redox with recent findings revealing that BLVRB alone can dictate cellular fate. However, as opposed to most enzymes, the BLVRB family remains enigmatic with an evolutionarily changing active site and unknown structural and functional consequences. Here, we applied a multi-faceted approach that combines X-ray crystallography, NMR and kinetics methods to elucidate the structural and functional basis of the evolutionarily changing BLVRB active site. Using a panel of three BLVRB isoforms (human, lemur and hyrax) and multiple human BLVRB mutants, our studies reveal a novel evolutionary mechanism where coenzyme 'clamps' formed by arginine side chains at two co-evolving positions within the active site serve to slow coenzyme release (Positions 14 and 78). We find that coenzyme release is further slowed by the weaker binding substrate, resulting in relatively slow turnover numbers. However, different BLVRB active sites imposed by either evolution or mutagenesis exhibit a surprising inverse relationship between coenzyme release and substrate turnover that is independent of the faster chemical step of hydride transfer also measured here. Collectively, our studies have elucidated the role of the evolutionarily changing BLVRB active site that serves to modulate coenzyme release and has revealed that coenzyme release is coupled to substrate turnover.

Keywords: biliverdin reductase B; coenzyme; dynamics; enzyme.

MeSH terms

  • Crystallography, X-Ray
  • Humans
  • Models, Molecular
  • Oxidoreductases Acting on CH-CH Group Donors / chemistry*
  • Oxidoreductases Acting on CH-CH Group Donors / isolation & purification
  • Oxidoreductases Acting on CH-CH Group Donors / metabolism*
  • Protein Conformation
  • Thermodynamics*

Substances

  • Oxidoreductases Acting on CH-CH Group Donors
  • biliverdin reductase