He was born at Constantinople of a distinguished family, and was a pupil of Gemistus Pletho. In 1390 he led an embassy sent to Venice by the emperor Manuel II Palaeologus to implore the aid of the Christian princes against the Turks. Roberto Rossi of Florence met him there, and in 1395 Rossi's acquaintance Jacopo Angeli da Scarperia set off for Constantinople to study Greek with Chrysoloras. In 1396 Coluccio Salutati, the chancellor of the University of Florence, invited him to come and teach Greek grammar and literature, quoting Cicero:
Having visited Milan and Pavia, and resided for several years at Venice, he went to Rome on the invitation of Bruni, who was then secretary to Pope Gregory XII. In 1408 be was sent to Paris on an important mission from the emperor Manuel Palaeologus. In 1413 he went to Germany on an embassy to the emperor Sigismund, the object of which was to fix a place for the church council that later assembled at Constance. Chrysoloras was on his way there, having been chosen to represent the Greek Church, when he died suddenly.
Though Chrysoloras became famous as a translator of Homer and Plato (The Republic), his works circulated in manuscript in his lifetime; two were eventually printed, his Erotemata (Questions). first published at Venice in 1484, and then widely reprinted, which was the first basic Greek grammar in use in Western Europe, and Epistolae III de comparatione veteris et novae Romae (Three Letters Comparing Ancient and Modern Rome). Many of his treatises on morals and ethics and other philosophical subjects came into print in the 17th and 18th centuries, because of their antiquarian interest.