Iron-sulfur clusters of biotin synthase in vivo: a Mössbauer study.
Biotin synthase, the enzyme that catalyzes the last step of the biosynthesis of biotin, contains only [2Fe-2S](2+) clusters when isolated under aerobic conditions. Previous results showed that reconstitution with an excess of FeCl(3) and Na(2)S under reducing and anaerobic conditions leads to either [4Fe-4S](2+), [4Fe-4S](+), or a mixture of [4Fe-4S](2+) and [2Fe-2S](2+) clusters. To determine whether any of these possibilities or other different cluster configuration could correspond to the physiological in vivo state, we have used (57)Fe Mössbauer spectroscopy to investigate the clusters of biotin synthase in whole cells. The results show that, in aerobically grown cells, biotin synthase contains a mixture of [4Fe-4S](2+) and [2Fe-2S](2+) clusters. A mixed [4Fe-4S](2+):[2Fe-2S](2+) cluster form has already been observed under certain in vitro conditions, and it has been proposed that both clusters might each play a significant role in the mechanism of biotin synthase. Their presence in vivo is now another argument in favor of this mixed cluster form.
Benda
Rüdiger
R
Institut für Physik, Universität zu Lübeck, Ratzeburger Allee 160, Germany.
Tse Sum Bui
Bernadette
B
Schünemann
Volker
V
Florentin
Dominique
D
Marquet
Andrée
A
Trautwein
Alfred X
AX
D000332:Aerobiosis; D002458:Cell Fractionation; D002498:Centrifugation; D004926:Escherichia coli / Q000201:enzymology* / Q000254:growth & development; D029968:Escherichia coli Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D007503:Iron Isotopes / Q000378:metabolism; D007506:Iron-Sulfur Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*; D013010:Sonication; D015204:Spectroscopy, Mossbauer / Q000379:methods; D013466:Sulfurtransferases / Q000096:biosynthesis / Q000302:isolation & purification / Q000378:metabolism*
D029968:Escherichia coli Proteins; D007503:Iron Isotopes; D007506:Iron-Sulfur Proteins; D013466:Sulfurtransferases; C057985:biotin synthetase