The aim of this study was to evaluate the effect of particle size distribution on composition, properties rheological, pasting, microstructural and baking properties of whole grain wheat flour (WGWF) of three different particles sizes (194.9 μm, 609.4 μm and 830.0 μm). The quantification of free sulfhydryl groups (-SH) of WGWF samples, together with the effects observed in the behavior of the dough and bread showed that particle size influences the functionality of the gluten network in a differentiated way. Firmer and lower breads volume compared to refined wheat flour (RF) were correlated with the quality of the gluten network. In the sample of finer particles, more pronounced adverse effects in quality (dough rheology, bread volume and texture) compared to the medium and coarse particle size sample suggests that the larger contact surface and the increased release of reactive compounds due to cell rupture interact with the gluten-forming proteins changing their functionality.