Thinopyrum intermedium, commonly known as intermediate wheatgrass (IWG) is a perennial crop that provides sustainable environmental benefits and shows a great potential to be developed as a grain crop [1]. Compared to wheat, IWG is generally higher in protein, fiber and antioxidants, but lower in total starch [2]. While higher in protein content, we found that the protein distribution/profile of IWG is considerably different than that of common wheat. The protein component in IWG consists mainly of gliadins and low molecular weight glutenins (LMWG), and is deficient in high molecular weight glutenins (HMWG), which are important for dough strength in bread applications.Fiber content in wheat flour negatively impacts gluten network formation and hence bread quality [3]. Fiber causes water redistribution in a dough system, thus altering the protein confirmation from spiral (consecutive turns) structure to sheet structure [4]. This main structural change in the gluten network due to fiber presence may result in physical disruption of gas cells producing poor quality bread. The fiber content is higher in whole IWG flour than in whole wheat flour due to higher ratio of bran to endosperm [5]. It is, therefore, hypothesized that the fiber interference would have a major impact on protein network formation in IWG dough system. With the fiber interference in mind, it is essential to optimize the mixing conditions (temperature and time) along with the flour refinement for desired protein network formation, as recently shown [6] for both strong and weak common wheat flours.Therefore, the objective of this study was to determine the effect of bran reduction along with various mixing conditions on protein network formation by monitoring changes in the protein's secondary structure and state of water in IWG dough using attenuated total reflectance (ATR) fourier transform infrared (FTIR) spectroscopy.IWG grains sample was milled a Quadrumat Junior flour mill (Brabender Inc.) and bran was separated. Bran was added back to refined IWG flour at 100%, 75%, 50%, 25% and 0% of original bran content. Different flour samples were evaluated for dough strength using farinograph following the constant flour weight procedure according to AACC method 54-21.02. Dough samples were collected at different time points during mixing: dough development time (DDT), stability departure, and over-mixing at two temperatures (21°C and 30°C). The infrared spectra of the dough samples were recorded using an ATR-FTIR spectrophotometer. Spectra of reference H2O/D2O mixtures, matched to the moisture content of the dough samples, were collected and used for subtraction of water contributions in the amide I region (1600-1700 cm -1 ) of the vector-normalized spectra. The quantitative estimation of protein secondary structure in the amide I region of dough was assigned as 1620-1644 cm -1 for β-sheets, 1644-1652 cm -1 for random coil, 1652-1660 cm -1 for α-helix, and 1660-1685 cm -1 for β-turn structures.Regardless of the mixing temperature, as the level o...