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- W141922969 abstract "The nifS gene product is a pyridoxal phosphate binding enzyme which catalyzes the desulfuration of L-cysteine to yield L-alanine and sulfur. This activity is required for the full activation of the two nitrogenase component proteins called Fe protein and MoFe protein. Because both Fe protein and MoFe protein have metalloclusters comprised of metals and inorganic sulfide, we have suggested that NIFS might supply the inorganic sulfide for Fe-S core formation. The mechanism for the desulfurization of L-cysteine catalyzed by NIFS was determined in several ways. The substrate analogs, L-allylglycine and L-vinylglycine, were shown to irreversibly inactivate NIFS by formation of a γ-methylcystathionyl or cystathionyl residue, respectively, through nucleophilic attack by an active site cysteinyl residue on the corresponding pyridoxal phosphate-analog adduct. The reactive cysteinyl residue, which is required for cysteine desulfurization activity, was identified as Cys325 by the specific alkylation of that residue and by site-directed mutagenesis experiments. Also, the formation of an enzyme-bound cysteinyl persulfide was identified as an intermediate in the NIFS-catalyzed reaction. Finally, evidence was obtained for an enamine intermediate in the formation of L-alanine. These results support a mechanism for NIFS-catalyzed desulfurization of L-cysteine which involves formation of a substrate cysteine-pyridoxal phosphate ketimine adduct and subsequent nucleophilic attack by the thiolate anion of Cys325 on the sulfur of the substrate cysteine. This leads to formation of a protein-bound persulfide, which is the proposed sulfur donor in Fe-S core formation, and a pyridoxal phosphate-bound enamine which is ultimately released as L-alanine. Using L-cysteine, ferrous ion, DTT, and MgATP, NIFS is able to catalyze the reactivation of the apo-Fe protein in vitro. The Fe4S4 cluster of the Fe protein reconstituted by NIFS is identical to the native one, as indicated by EPR analysis in the presence or absence of MgATP." @default.
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- W141922969 date "1994-01-01" @default.
- W141922969 modified "2023-09-23" @default.
- W141922969 title "Involvement of PLP in biological formation iron-sulfur clusters" @default.
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- W141922969 doi "https://doi.org/10.1007/978-3-0348-7393-2_34" @default.
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