Crystal Structure of the Emerging Cancer Target MTHFD2 in Complex with a Substrate-Based Inhibitor

Research output: Contribution to journalArticle


To sustain their proliferation, cancer cells become dependent on one-carbon metabolism to support purine and thymidylate synthesis. Indeed, one of the most highly upregulated enzymes during neoplastic transformation is MTHFD2, a mitochondrial methylenetetrahydrofolate dehydrogenase and cyclohydrolase involved in one-carbon metabolism. Because MTHFD2 is expressed normally only during embryonic development, it offers a disease-selective therapeutic target for eradicating cancer cells while sparing healthy cells. Here we report the synthesis and preclinical characterization of the first inhibitor of human MTHFD2. We also disclose the first crystal structure of MTHFD2 in complex with a substrate-based inhibitor and the enzyme cofactors NAD+ and inorganic phosphate. Our work provides a rationale for continued development of a structural framework for the generation of potent and selective MTHFD2 inhibitors for cancer treatment. Cancer Res; 77(4); 937-48. ©2017 AACR.


  • Robert Gustafsson
  • Ann-Sofie Jemth
  • Nina M S Gustafsson
  • Katarina Färnegårdh
  • Olga Loseva
  • Elisée Wiita
  • Nadilly Bonagas
  • Leif Dahllund
  • Sabin Llona-Minguez
  • Maria Häggblad
  • Martin Henriksson
  • Yasmin Andersson
  • Evert Homan
  • Thomas Helleday
  • Pål Stenmark
External organisations
  • Stockholm University
  • Karolinska Institutet
Research areas and keywords


  • Binding Sites, Crystallization, Enzyme Inhibitors/chemistry, Folic Acid/analogs & derivatives, Humans, Leucovorin/analogs & derivatives, Methenyltetrahydrofolate Cyclohydrolase/antagonists & inhibitors, Methylenetetrahydrofolate Dehydrogenase (NADP)/antagonists & inhibitors, Minor Histocompatibility Antigens, Mitochondria/enzymology, NAD/metabolism, Protein Multimerization
Original languageEnglish
Pages (from-to)937-948
JournalCancer Research
Issue number4
Publication statusPublished - 2017 Feb 15
Publication categoryResearch
Externally publishedYes

Bibliographic note

©2016 American Association for Cancer Research.