The influences of egg white (EW), egg yolk (EY) and whole egg (WE) on the structure of highland barley dough and the quality of highland barley bread were explored. The results showed that egg powder reduced G' and G" of highland barley dough, which led to the softer texture of dough and endowed bread with a larger specific volume. EW increased the percentage of β-sheet of highland barley dough, EY and WE promoted the transformation from random coil to β-sheet and α-helix. Meanwhile, more disulfide bonds were formed from free sulfhydryl groups in the doughs with EY and WE. These properties of highland barley dough could help highland barley bread develop a preferable appearance and textural feature. It is worth noting that highland barley bread containing EY has more flavorful substances and a better crumb structure, which were similar to that of whole wheat bread. The highland barley bread with EY received a high score according to the sensory evaluation in consumer acceptance.