Crystal structure of Schizosaccharomyces pombe riboflavin kinase reveals a novel ATP and riboflavin-binding fold

J Mol Biol. 2003 Mar 7;326(5):1463-73. doi: 10.1016/s0022-2836(03)00059-7.

Abstract

The essential redox cofactors riboflavin monophosphate (FMN) and flavin adenine dinucleotide (FAD) are synthesised from their precursor, riboflavin, in sequential reactions by the metal-dependent riboflavin kinase and FAD synthetase. Here, we describe the 1.6A crystal structure of the Schizosaccharomyces pombe riboflavin kinase. The enzyme represents a novel family of phosphoryl transferring enzymes. It is a monomer comprising a central beta-barrel clasped on one side by two C-terminal helices that display an L-like shape. The opposite side of the beta-barrel serves as a platform for substrate binding as demonstrated by complexes with ADP and FMN. Formation of the ATP-binding site requires significant rearrangements in a short alpha-helix as compared to the substrate free form. The diphosphate moiety of ADP is covered by the glycine-rich flap I formed from parts of this alpha-helix. In contrast, no significant changes are observed upon binding of riboflavin. The ribityl side-chain might be covered by a rather flexible flap II. The unusual metal-binding site involves, in addition to the ADP phosphates, only the strictly conserved Thr45. This may explain the preference for zinc observed in vitro.

MeSH terms

  • Adenosine Diphosphate / chemistry
  • Adenosine Diphosphate / metabolism*
  • Adenosine Triphosphate / chemistry
  • Adenosine Triphosphate / metabolism*
  • Amino Acid Sequence
  • Binding Sites
  • Crystallization
  • Crystallography, X-Ray
  • Flavin Mononucleotide / chemistry
  • Flavin Mononucleotide / metabolism*
  • In Vitro Techniques
  • Models, Molecular
  • Molecular Sequence Data
  • Multienzyme Complexes / chemistry*
  • Multienzyme Complexes / metabolism
  • Phosphotransferases (Alcohol Group Acceptor) / chemistry*
  • Phosphotransferases (Alcohol Group Acceptor) / metabolism
  • Protein Binding
  • Protein Conformation
  • Protein Folding
  • Riboflavin / chemistry*
  • Riboflavin / metabolism
  • Schizosaccharomyces / enzymology*
  • Schizosaccharomyces pombe Proteins / chemistry*
  • Schizosaccharomyces pombe Proteins / metabolism
  • Sequence Homology, Amino Acid
  • Substrate Specificity

Substances

  • Multienzyme Complexes
  • Schizosaccharomyces pombe Proteins
  • Adenosine Diphosphate
  • Flavin Mononucleotide
  • Adenosine Triphosphate
  • Phosphotransferases (Alcohol Group Acceptor)
  • riboflavin kinase
  • Riboflavin

Associated data

  • PDB/1N05
  • PDB/1N06
  • PDB/1N07
  • PDB/1N08