Glutenin polymers composed of HMW and LMW subunits are important contributors to the wheat end-use properties. 26 winter wheat cultivars differing in bread processing quality were collected at the experimental fields of the Agricultural Institute Osijek, Croatia and Institute of the Field and Vegetable Crops Novi Sad, Serbia, in 2008/2009 season. The HMW glutenins composition and glutenin proteins content were determined by SDS-PAGE and RP-HPLC, respectively, with aim to determine the relationship between glutenin protein fractions and wheat quality properties. Significant differences were found between Croatian and Serbian cultivars in several quality attributes (GI, WA, DDT, DS and R/EXT) as well as in the content of total glutenins and LMW glutenins and GLI/GLU ratio. The dominant HMW subunits in analyzed cultivars were 2*, 7+9/7+8 and 5+10. Principal component analysis (PCA) confirmed the presence of association between HMW glutenins composition and GI, dough E, R and R/EXT, while the glutenins quantitative data showed pronounced relation with P, DDT, DS, E, R and R/EXT. GLI/GLU ratio had the opposite effect on these parameters.