Protein extraction from wheat bran is challenging due to its multi-layer and fiber-rich structure. Here, opening aleurone cells, via dry and wet milling, their combination and a novel ultrafine milling, and its effect on wheat bran’s protein recovery using the alkaline solubilization/isoelectric precipitation and protein structure, functionality, and phytate content were investigated. Wet milling and ultrafine milling improved protein recovery and purity but only ultrafine milling reduced bran particle size to the aleurone cells and exposed their structure. Despite this, ultrafine milling did not significantly increase protein yield compared to wet milling, which partially opened the aleurone cells, meaning that opening the cells per se is not enough for extracting their protein. Proteins extracted with the aid of ultrafine milling had smaller particle sizes with significantly better water solubility (>2-fold) and rheological properties. Both wet milling and ultrafine milling significantly improved the removal of phytate during the wet fractionation process. Altogether, optimizing milling techniques offers a promising path to enhance accessibility to wheat bran proteins and their quality if carefully fine-tuned but other assistant technologies are necessary for boosting the recovery of the released protein from aleurone cells.