Cardiolipin (CL) is an evolutionarily conserved phospholipid which exists almost exclusively in the inner mitochondrial membrane (IMM) of mammalian cells and plasma membranes of bacteria. This phospholipid plays an important role in mitochondrial bioenergetics by affecting the activity of key proteins of IMM. The structure of CL is characterized by a glycerol backbone connected to two phosphatidyl lipids. Two chiral carbons and four fatty acyl chains in CL result in a flexible body allowing interactions with respiratory chain complexes and mitochondrial substrate carriers. The biosynthesis of CL is a little more complex than that of phosphatidylethanolamine (PE) and phosphatidylserine (PS), and it involves two pathways: de novo synthesis and remodeling steps.