He studied at the School of Arts (1902- 1906) under his father Nikephoros Lytras and Georgios Jakovides. In 1907 he continued his studies at the Munich Academy under Ludwig von Lofftz. In the Bavarian capital he came into contact with German expressionism and the creations of the Blue Rider group. After his return to Athens in 1912 he began to participate in the exhibitions of the League of Greek Artists (1915, 1916, 1917, 1920, 1926) while in 1919 he exhibited in tandem with the sculptor Grigorios Zevgolis. He was a founding member of the Art Group. In 1923 he was elected professor at the School of Fine Arts. After his death, retrospectives of his work were organized in 1929 at the Zappeion Hall and in 1936 at the Venice Biennale.

Nikolaos Lytras, Konstantinos Parthenis and Konstantinos Maleas are considered to have revitalized modern Greek art at the beginning of the 20th century. He influenced the development of Greek painting both through his teaching at the School of Fine Arts and his pioneering art work. Portraits, landscapes and still lifes were built up with broad, free brushstrokes of viscous color and gestural writing.