The improvement of the processing properties of gluten-free cereals is a challenging issue for cereal technologists. Vital wheat gluten (VWG), transglutaminase (TGase), or combinations of the two have been successfully used to improve the processing properties of food materials. In this study, a Mixolab system and a rheometer were used to investigate the effects of TGase on the thermomechanical and dynamic rheological properties of buckwheat flour dough (BF dough) and a buckwheat flour dough containing 15% VWG (BF-VWG dough). The results indicated that the addition of 1.0% TGase decreased the water absorption and significantly increased the stability, storage modulus (G′), and complex modulus (|G*|) of the BF dough. The presence of 15% VWG had a synergistic effect, enhancing the effectiveness of the TGase in improving the thermomechanical and dynamic rheological properties of the BF-VWG dough. Quantification of the free amino groups showed that the amino groups were more effectively bonded in the BF-VWG dough than in the BF dough when the TGase level exceeded 1.0%. Confocal laser scanning microscopy (CLSM) and sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis confirmed that the TGase promoted the formation of protein complexes. These data indicate that the combination of VWG and TGase can be used to improve the properties of buckwheat flour for buckwheat-based food production.