铁硫蛋白（英语：Iron-sulfur protein，或称为铁硫蛋白质）是一类蛋白质，其特征是其中存在着铁-硫簇，铁-硫簇中含有与硫连接着的二、三或四个铁中心，并可处于各种变化的氧化态上。铁-硫簇存在于多种金属蛋白之中，例如铁氧还蛋白、氢化酶、辅酶Q-细胞色素c氧化酶、琥珀酸脱氢酶，和固氮酶等。铁硫簇是粒线体中电子传递途径中的重要成员，像氧化磷酸化过程中的第一蛋白复合体（Complex I）及第二蛋白复合体（Complex II）都含有铁硫簇。此外，像是顺乌头酸酶和SAM依赖蛋白中也有铁硫簇，另外硫辛酸及生物素的合成也牵涉到铁硫簇。可见铁硫簇的作用范围包括呼吸作用、光合作用、羟化作用以及细菌的氢和氮的固定。

Iron–sulfur proteins are proteins characterized by the presence of iron–sulfur clusters containing sulfide-linked di-, tri-, and tetrairon centers in variable oxidation states. Iron–sulfur clusters are found in a variety of metalloproteins, such as the ferredoxins, as well as NADH dehydrogenase, hydrogenases, coenzyme Q – cytochrome c reductase, succinate – coenzyme Q reductase and nitrogenase. Iron–sulfur clusters are best known for their role in the oxidation-reduction reactions of mitochondrial electron transport. Both Complex I and Complex II of oxidative phosphorylation have multiple Fe–S clusters. They have many other functions including catalysis as illustrated by aconitase, generation of radicals as illustrated by SAM-dependent enzymes, and as sulfur donors in the biosynthesis of lipoic acid and biotin. Additionally, some Fe–S proteins regulate gene expression. Fe–S proteins are vulnerable to attack by biogenic nitric oxide. In most Fe–S proteins, the terminal ligands on Fe are thiolate, but exceptions exist.