There is evidence suggesting that transglutaminase (TG) 2 plays a role in stabilizing monomeric and aggregated huntingtin, thereby contributing to the pathophysiology of Huntington disease. Calmodulin (CaM) regulates TG2 cross-linking of N-terminal mutant huntingtin in cells and colocalizes with TG and huntingtin in inclusions in Huntington disease cortex. The current study examined the effects of small fragments of CaM in human embryonic kidney 293T cells expressing N-terminal mutant huntingtin and transglutaminase 2. Four CaM fragments were developed: first 76 amino acids, last 72 amino acids, 77 amino acids in the center (CaM-center), and the overlapping region of last 72 amino acids and CaM-center (CaM-overlap). The last 72 amino acids, CaM-center, and CaM-overlap significantly decreased amounts of TG-modified huntingtin by 40% to 60%, and cytotoxicity decreased up to 40% compared with cells not expressing any CaM construct. Carbachol-stimulated release of intracellular calcium is significantly higher in cells expressing N-terminal mutant huntingtin and TG2 compared with vector-transfected cells; expression of either CaM-center or CaM-overlap in these cells returned the levels of carbachol-stimulated intracellular calcium release to control values. Furthermore, CaM-overlap expression significantly decreased huntingtin binding to CaM. These data further suggest that CaM regulates TG2 activity, plays a role in the disease-related modifications to mutant huntingtin, and that disruption of CaM-huntingtin interaction is potentially a new target for therapeutic intervention in Huntington disease.