Berruguete is believed to have studied under Fernando Gallego or Colantonio and to have worked about 1474 at the “studiolo” of Federico da Montefeltro in the ducal palace in Urbino (Italy). He returned to Spain to produce works for the Toledo Cathedral (1483–99) and then moved to Ávila to work on the cathedral there until his death.

Berruguete's paintings are marked by elegance and dignity combined with a profound feeling for space and atmosphere. The style is often eclectic, a mixture of Flemish, Spanish Gothic, and Italian Renaissance, but it is never imitative. This combination of influences may be seen in his masterpiece, the main altarpiece in the convent of San Tomás in Ávila (1499–1503). In these panels, representing scenes from the life of St. Thomas of Ávila, there is a combination of luxurious ornament and gold decoration with a simplicity and ease of depiction.