Aragon also previously controlled the Duchy of Milan, but a year before Charles ascended to the throne, it was annexed by France after the Battle of Marignano in 1515.
The Knight of Malta were part of the Holy League
Barbarossa's naval victories secured Ottoman dominance over the Mediterranean during the mid 16th century, from the Battle of Preveza in 1538 until the Battle of Lepanto in 1571.
Andrea Doria was the losing admiral at the Battle of Preveza
Pope Paul III put forth the call for a Holy League against the Ottomans
The Republic of Venice urged Pope Paul III to assemble a Holy League against the Ottoman Empire
Hapsburg Empire during the reign of Charles V
Charles succeeded in re-capturing Milan in 1522 when Imperial troops defeated the Franco-Swiss army at Bicocca.
Francis I was taken at the Battle of Pavia in 1526