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In biochemistry, the iron-sulfur cluster biosynthesis describes the components and processes involved in the biosynthesis of iron-sulfur proteins. The topic is of interest because these proteins are pervasive. The iron sulfur proteins contain iron-sulfur clusters, some with elaborate structures, that feature iron and sulfide centers. One broad biosynthetic task is producing sulfide , which requires various families of enzymes. Another broad task is affixing the sulfide to iron, which is achieved on scaffolds, which are nonfunctional. Finally these Fe-S cluster is transferred to a target protein, which then become functional.

The formation of iron-sulfur clusters are produced by one of four pathways:

In addition to those three systems, the so-called Cystosolic Iron-Sulfur Assembly is invoked for cytosolic and nuclear Fe–S proteins.

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