Introduction of a [4Fe-4S (S-cys)4]+1,+2 iron-sulfur center into a four-α helix protein using design parameters from the domain of the Fx cluster in the Photosystem I reaction center

dc.contributor.author Scott, Marvin
dc.contributor.author Biggins, John
dc.contributor.author Scott, Marvin
dc.contributor.department Agronomy
dc.date 2018-02-17T13:45:12.000
dc.date.accessioned 2020-06-29T23:07:02Z
dc.date.available 2020-06-29T23:07:02Z
dc.date.issued 1997-02-01
dc.description.abstract <p>We describe the insertion of an iron-sulfur center into a designed four α-helix model protein. The model protein was re-engineered by introducing four cysteine ligands required for the coordination of the mulinucleate cluster into positions in the main-chain directly analogous to the domain predicted to ligand the interpeptide [4Fe-4S (S-cys)4] cluster, Fx, from PsaA and PsaB of the Photosystem I reaction center. This was achieved by inserting the sequence, CDGPGRGGTC, which is conserved in PsaA and PsaB, into interhelical loops 1 and 3 of the four α-helix model. The holoprotein was characterized spectroscopically after insertion of the iron-sulfur center in vitro. EPR spectra confirmed the cluster is a [4Fe-4S] type, indicating that the cysteine thiolate ligands were positioned as designed. The midpoint potential of the iron-sulfur center in the model holoprotein was determined via redox titration and shown to be −422 mV (pH 8.3,<em>n</em> = 1). The results support proposals advanced for the structure of the domain of the [4Fe-4S] Fx cluster in Photosystem I based upon sequence predictions and molecular modeling. We suggest that the lower potential of the Fx cluster is most likely due to factors in the protein environment of Fx rather than the identity of the residues proximal to the coordinating ligands.</p>
dc.description.comments <p>This article is from <em>Protein Science</em> 6 (1997): 340, doi:<a href="http://dx.doi.org/10.1002/pro.5560060209" target="_blank">10.1002/pro.5560060209</a>. </p>
dc.format.mimetype application/pdf
dc.identifier archive/lib.dr.iastate.edu/agron_pubs/78/
dc.identifier.articleid 1088
dc.identifier.contextkey 8192609
dc.identifier.s3bucket isulib-bepress-aws-west
dc.identifier.submissionpath agron_pubs/78
dc.identifier.uri https://dr.lib.iastate.edu/handle/20.500.12876/5050
dc.language.iso en
dc.source.bitstream archive/lib.dr.iastate.edu/agron_pubs/78/1997_ScottMP_Introduction4Fe4S.pdf|||Sat Jan 15 01:54:06 UTC 2022
dc.source.uri 10.1002/pro.5560060209
dc.subject.disciplines Agronomy and Crop Sciences
dc.subject.disciplines Biochemistry
dc.subject.disciplines Cell Biology
dc.subject.disciplines Molecular Biology
dc.subject.keywords four-helix bundle protein
dc.subject.keywords iron-sulfur center
dc.subject.keywords Photosystem I reaction center
dc.subject.keywords protein design
dc.title Introduction of a [4Fe-4S (S-cys)4]+1,+2 iron-sulfur center into a four-α helix protein using design parameters from the domain of the Fx cluster in the Photosystem I reaction center
dc.type article
dc.type.genre article
dspace.entity.type Publication
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relation.isOrgUnitOfPublication fdd5c06c-bdbe-469c-a38e-51e664fece7a
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