• GFOD1 lacks cofactor binding and catalytic residues, confirming its role as a pseudoenzyme in the Gfo/Idh/MocA family.
• Crystal structure reveals GFOD1 forms a homodimer, yet is structurally distinct from active oxidoreductases.
• GFOD1 interacts with both GTP- and GDP-bound NKIRAS2, suggesting a regulatory role in NF-κB signaling.
• The GFOD1-NKIRAS2 interaction is mediated by the interswitch region of NKIRAS2, providing a potential therapeutic target for psychiatric and inflammatory disorders.