The effects of transglutaminase treatment (0–2 units/g milk protein) on the chemical composition, textural characteristics, proteolysis and yield of reduced-fat Iranian white cheese (milk fat: 0.4–1.4% w/w) incorporated with whey proteins (0–6 g/L milk) were investigated. Enzyme-mediated inclusion of whey proteins in the reduced-fat cheese caused a noticeable increase in moisture to protein (M:P) ratio with concomitant decease in the hardness rheological parameters of fracture stress and Young’s and storage (G’) moduli. However, increase in concentrations of whey proteins or/and transglutaminase enzyme above a critical level led to formation of a cheese matrix with lower moisture content and greater values of hardness indices. Whey protein addition and transglutaminase treatment resulted in the same trends of changes in proteolysis rate and cheese yield as in cheese softness. Response surface method (RSM) suggested that the enzymatic incorporation of 4.2 g deliberately added whey proteins to 1 L of milk (1.04% w/w fat) into the cheese matrix using 0.833 unit transglutaminase per gram milk protein would provide a reduced-fat product with the softest texture and the highest yield. The scanning electron micrographs showed formation of honeycomb structures in the protein matrix of the reduced-fat sample with optimum formulation.