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The primary conduit is MtrCAB, a trimeric porin\u2010cytochrome complex that contains the cell surface exposed decaheme cytochrome MtrC. This donates electrons to extracellular substrates, including OmcA, soluble metals, organic electron shuttles, and insoluble metal oxides. However, it is not clear whether this broad substrate specificity requires specific sites for binding and reduction, or whether reduction occurs through non\u2010specific interactions near exposed hemes on the cytochrome surface. <jats:italic>Shewanella oneidensis<\/jats:italic> MtrC is composed of four domains, with the hemes closely packed and distributed evenly between domains II and IV. The domains are arranged to allow electron transport across the cytochrome via interdomain electron transfer, but the significance of this conserved feature is not understood. Here we use site\u2010directed mutagenesis to generate an MtrC variant that is comprised only of domains I and II (MtrC<jats:sub>DI,II<\/jats:sub>). The properties of this MtrC<jats:sub>DI,II<\/jats:sub> are effectively identical to domains I and II of full\u2010length MtrC. Whole\u2010cell assays revealed that <jats:italic>S. oneidensis<\/jats:italic> cells replacing full\u2010length MtrC with MtrC<jats:sub>DI,II<\/jats:sub> had significantly lower rates of OmcA, flavin mononucleotide, and Fe(III) citrate reduction. Our results demonstrate that MtrC domains III and IV contain sites for association of specific substrates, enabling the reduction of extracellular electron acceptors in <jats:italic>S. oneidensis<\/jats:italic>.<\/jats:p>","DOI":"10.1002\/pro.70243","type":"journal-article","created":{"date-parts":[[2025,7,29]],"date-time":"2025-07-29T05:03:38Z","timestamp":1753765418000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["Extracellular catalysis of environmental substrates by <i>Shewanella oneidensis<\/i><scp>MR<\/scp>\u20101 occurs via active sites on the C\u2010terminal domains of <scp>MtrC<\/scp>"],"prefix":"10.1002","volume":"34","author":[{"ORCID":"https:\/\/orcid.org\/0009-0007-0938-0790","authenticated-orcid":false,"given":"Alejandro","family":"Morales\u2010Florez","sequence":"first","affiliation":[{"name":"School of Biological Sciences University of East Anglia  Norwich UK"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Colin W. 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