The high-resolution crystal structure of periplasmic Haemophilus influenzae NAD nucleotidase reveals a novel enzymatic function of human CD73 related to NAD metabolism

Biochem J. 2012 Jan 1;441(1):131-41. doi: 10.1042/BJ20111263.

Abstract

Haemophilus influenzae is a major pathogen of the respiratory tract in humans that has developed the capability to exploit host NAD(P) for its nicotinamide dinucleotide requirement. This strategy is organized around a periplasmic enzyme termed NadN (NAD nucleotidase), which plays a central role by degrading NAD into adenosine and NR (nicotinamide riboside), the latter being subsequently internalized by a specific permease. We performed a biochemical and structural investigation on H. influenzae NadN which determined that the enzyme is a Zn2+-dependent 5'-nucleotidase also endowed with NAD(P) pyrophosphatase activity. A 1.3 Å resolution structural analysis revealed a remarkable conformational change that occurs during catalysis between the open and closed forms of the enzyme. NadN showed a broad substrate specificity, recognizing either mono- or di-nucleotide nicotinamides and different adenosine phosphates with a maximal activity on 5'-adenosine monophosphate. Sequence and structural analysis of H. influenzae NadN led us to discover that human CD73 is capable of processing both NAD and NMN, therefore disclosing a possible novel function of human CD73 in systemic NAD metabolism. Our data may prove to be useful for inhibitor design and disclosed unanticipated fascinating evolutionary relationships.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 5'-Nucleotidase / metabolism*
  • Adenosine Diphosphate
  • Amino Acid Sequence
  • Animals
  • Bacterial Proteins / genetics
  • Bacterial Proteins / metabolism*
  • Binding Sites
  • COS Cells
  • Chlorocebus aethiops
  • Cloning, Molecular
  • Crystallization
  • Gene Expression Regulation, Bacterial / physiology*
  • Haemophilus influenzae / enzymology*
  • Haemophilus influenzae / genetics
  • Haemophilus influenzae / metabolism
  • Humans
  • Models, Molecular
  • Molecular Sequence Data
  • NAD / metabolism
  • Nicotinamide Mononucleotide / metabolism
  • Nucleotidases / genetics
  • Nucleotidases / metabolism*
  • Protein Conformation
  • Pyrophosphatases / genetics
  • Pyrophosphatases / metabolism*
  • Zinc / chemistry

Substances

  • Bacterial Proteins
  • NAD
  • Nicotinamide Mononucleotide
  • Adenosine Diphosphate
  • Nucleotidases
  • 5'-Nucleotidase
  • NadN protein, Haemophilus influenzae
  • Pyrophosphatases
  • Zinc