Characterization of the Cofactor Composition of Escherichia coli Biotin Synthase

Michele Mader Cosper, Guy N.L. Jameson, Heather L. Hernández, Carsten Krebs, Boi Hanh Huynh, Michael K. Johnson

Research output: Contribution to journalArticle

79 Citations (Scopus)

Abstract

The cofactor content of in vivo, as-isolated, and reconstituted forms of recombinant Escherichia coli biotin synthase (BioB) has been investigated using the combination of UV-visible absorption, resonance Raman, and Mössbauer spectroscopies along with parallel analytical and activity assays. In contrast to the recent report that E. coli BioB is a pyridoxal phosphate (PLP)-dependent enzyme with intrinsic cysteine desulfurase activity (Ollagnier-deChoudens, S., Mulliez, E., Hewitson, K. S., and Fontecave, M. (2002) Biochemistry 41, 9145-9152), no evidence for PLP binding or for PLP-induced cysteine desulfurase or biotin synthase activity was observed with any of the forms of BioB investigated in this work. The results confirm that BioB contains two distinct Fe-S cluster binding sites. One site accommodates a [2Fe-2S]2+ cluster with partial noncysteinyl ligation that can only be reconstituted in vitro in the presence of O2. The other site accommodates a [4Fe-4S]2+,+ cluster that binds S-adenosylmethionine (SAM) at a unique Fe site of the [4Fe-4S]2+ cluster and undergoes O 2-induced degradation via a distinct type of [2Fe-2S]2+ cluster intermediate. In vivo Mössbauer studies show that recombinant BioB in anaerobically grown cells is expressed exclusively in an inactive form containing only the as-isolated [2Fe-2S]2+ cluster and demonstrate that the [2Fe-2S]2+ cluster is not a consequence of overexpressing the recombinant enzyme under aerobic growth conditions. Overall the results clarify the confusion in the literature concerning the Fe-S cluster composition and the in vitro reconstitution and O2-induced cluster transformations that are possible for recombinant BioB. In addition, they provide a firm foundation for assessing cluster transformations that occur during turnover and the catalytic competence of the [2Fe-2S]2+ cluster as the immediate S-donor for biotin biosynthesis.

Original languageEnglish (US)
Pages (from-to)2007-2021
Number of pages15
JournalBiochemistry
Volume43
Issue number7
DOIs
StatePublished - Feb 24 2004

Fingerprint

Pyridoxal Phosphate
Escherichia coli
Cysteine Synthase
Chemical analysis
S-Adenosylmethionine
Biochemistry
Raman Spectrum Analysis
Biosynthesis
Enzymes
Biotin
Mental Competency
Ligation
Assays
Binding Sites
Spectroscopy
Degradation
Growth
biotin synthetase
cysteine desulfurase
In Vitro Techniques

All Science Journal Classification (ASJC) codes

  • Biochemistry

Cite this

Cosper, M. M., Jameson, G. N. L., Hernández, H. L., Krebs, C., Huynh, B. H., & Johnson, M. K. (2004). Characterization of the Cofactor Composition of Escherichia coli Biotin Synthase. Biochemistry, 43(7), 2007-2021. https://doi.org/10.1021/bi0356653
Cosper, Michele Mader ; Jameson, Guy N.L. ; Hernández, Heather L. ; Krebs, Carsten ; Huynh, Boi Hanh ; Johnson, Michael K. / Characterization of the Cofactor Composition of Escherichia coli Biotin Synthase. In: Biochemistry. 2004 ; Vol. 43, No. 7. pp. 2007-2021.
@article{2aa18ed1f0e746bc8ac3ed248d1221c9,
title = "Characterization of the Cofactor Composition of Escherichia coli Biotin Synthase",
abstract = "The cofactor content of in vivo, as-isolated, and reconstituted forms of recombinant Escherichia coli biotin synthase (BioB) has been investigated using the combination of UV-visible absorption, resonance Raman, and M{\"o}ssbauer spectroscopies along with parallel analytical and activity assays. In contrast to the recent report that E. coli BioB is a pyridoxal phosphate (PLP)-dependent enzyme with intrinsic cysteine desulfurase activity (Ollagnier-deChoudens, S., Mulliez, E., Hewitson, K. S., and Fontecave, M. (2002) Biochemistry 41, 9145-9152), no evidence for PLP binding or for PLP-induced cysteine desulfurase or biotin synthase activity was observed with any of the forms of BioB investigated in this work. The results confirm that BioB contains two distinct Fe-S cluster binding sites. One site accommodates a [2Fe-2S]2+ cluster with partial noncysteinyl ligation that can only be reconstituted in vitro in the presence of O2. The other site accommodates a [4Fe-4S]2+,+ cluster that binds S-adenosylmethionine (SAM) at a unique Fe site of the [4Fe-4S]2+ cluster and undergoes O 2-induced degradation via a distinct type of [2Fe-2S]2+ cluster intermediate. In vivo M{\"o}ssbauer studies show that recombinant BioB in anaerobically grown cells is expressed exclusively in an inactive form containing only the as-isolated [2Fe-2S]2+ cluster and demonstrate that the [2Fe-2S]2+ cluster is not a consequence of overexpressing the recombinant enzyme under aerobic growth conditions. Overall the results clarify the confusion in the literature concerning the Fe-S cluster composition and the in vitro reconstitution and O2-induced cluster transformations that are possible for recombinant BioB. In addition, they provide a firm foundation for assessing cluster transformations that occur during turnover and the catalytic competence of the [2Fe-2S]2+ cluster as the immediate S-donor for biotin biosynthesis.",
author = "Cosper, {Michele Mader} and Jameson, {Guy N.L.} and Hern{\'a}ndez, {Heather L.} and Carsten Krebs and Huynh, {Boi Hanh} and Johnson, {Michael K.}",
year = "2004",
month = "2",
day = "24",
doi = "10.1021/bi0356653",
language = "English (US)",
volume = "43",
pages = "2007--2021",
journal = "Biochemistry",
issn = "0006-2960",
publisher = "American Chemical Society",
number = "7",

}

Cosper, MM, Jameson, GNL, Hernández, HL, Krebs, C, Huynh, BH & Johnson, MK 2004, 'Characterization of the Cofactor Composition of Escherichia coli Biotin Synthase', Biochemistry, vol. 43, no. 7, pp. 2007-2021. https://doi.org/10.1021/bi0356653

Characterization of the Cofactor Composition of Escherichia coli Biotin Synthase. / Cosper, Michele Mader; Jameson, Guy N.L.; Hernández, Heather L.; Krebs, Carsten; Huynh, Boi Hanh; Johnson, Michael K.

In: Biochemistry, Vol. 43, No. 7, 24.02.2004, p. 2007-2021.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Characterization of the Cofactor Composition of Escherichia coli Biotin Synthase

AU - Cosper, Michele Mader

AU - Jameson, Guy N.L.

AU - Hernández, Heather L.

AU - Krebs, Carsten

AU - Huynh, Boi Hanh

AU - Johnson, Michael K.

PY - 2004/2/24

Y1 - 2004/2/24

N2 - The cofactor content of in vivo, as-isolated, and reconstituted forms of recombinant Escherichia coli biotin synthase (BioB) has been investigated using the combination of UV-visible absorption, resonance Raman, and Mössbauer spectroscopies along with parallel analytical and activity assays. In contrast to the recent report that E. coli BioB is a pyridoxal phosphate (PLP)-dependent enzyme with intrinsic cysteine desulfurase activity (Ollagnier-deChoudens, S., Mulliez, E., Hewitson, K. S., and Fontecave, M. (2002) Biochemistry 41, 9145-9152), no evidence for PLP binding or for PLP-induced cysteine desulfurase or biotin synthase activity was observed with any of the forms of BioB investigated in this work. The results confirm that BioB contains two distinct Fe-S cluster binding sites. One site accommodates a [2Fe-2S]2+ cluster with partial noncysteinyl ligation that can only be reconstituted in vitro in the presence of O2. The other site accommodates a [4Fe-4S]2+,+ cluster that binds S-adenosylmethionine (SAM) at a unique Fe site of the [4Fe-4S]2+ cluster and undergoes O 2-induced degradation via a distinct type of [2Fe-2S]2+ cluster intermediate. In vivo Mössbauer studies show that recombinant BioB in anaerobically grown cells is expressed exclusively in an inactive form containing only the as-isolated [2Fe-2S]2+ cluster and demonstrate that the [2Fe-2S]2+ cluster is not a consequence of overexpressing the recombinant enzyme under aerobic growth conditions. Overall the results clarify the confusion in the literature concerning the Fe-S cluster composition and the in vitro reconstitution and O2-induced cluster transformations that are possible for recombinant BioB. In addition, they provide a firm foundation for assessing cluster transformations that occur during turnover and the catalytic competence of the [2Fe-2S]2+ cluster as the immediate S-donor for biotin biosynthesis.

AB - The cofactor content of in vivo, as-isolated, and reconstituted forms of recombinant Escherichia coli biotin synthase (BioB) has been investigated using the combination of UV-visible absorption, resonance Raman, and Mössbauer spectroscopies along with parallel analytical and activity assays. In contrast to the recent report that E. coli BioB is a pyridoxal phosphate (PLP)-dependent enzyme with intrinsic cysteine desulfurase activity (Ollagnier-deChoudens, S., Mulliez, E., Hewitson, K. S., and Fontecave, M. (2002) Biochemistry 41, 9145-9152), no evidence for PLP binding or for PLP-induced cysteine desulfurase or biotin synthase activity was observed with any of the forms of BioB investigated in this work. The results confirm that BioB contains two distinct Fe-S cluster binding sites. One site accommodates a [2Fe-2S]2+ cluster with partial noncysteinyl ligation that can only be reconstituted in vitro in the presence of O2. The other site accommodates a [4Fe-4S]2+,+ cluster that binds S-adenosylmethionine (SAM) at a unique Fe site of the [4Fe-4S]2+ cluster and undergoes O 2-induced degradation via a distinct type of [2Fe-2S]2+ cluster intermediate. In vivo Mössbauer studies show that recombinant BioB in anaerobically grown cells is expressed exclusively in an inactive form containing only the as-isolated [2Fe-2S]2+ cluster and demonstrate that the [2Fe-2S]2+ cluster is not a consequence of overexpressing the recombinant enzyme under aerobic growth conditions. Overall the results clarify the confusion in the literature concerning the Fe-S cluster composition and the in vitro reconstitution and O2-induced cluster transformations that are possible for recombinant BioB. In addition, they provide a firm foundation for assessing cluster transformations that occur during turnover and the catalytic competence of the [2Fe-2S]2+ cluster as the immediate S-donor for biotin biosynthesis.

UR - http://www.scopus.com/inward/record.url?scp=1242285458&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=1242285458&partnerID=8YFLogxK

U2 - 10.1021/bi0356653

DO - 10.1021/bi0356653

M3 - Article

C2 - 14967041

AN - SCOPUS:1242285458

VL - 43

SP - 2007

EP - 2021

JO - Biochemistry

JF - Biochemistry

SN - 0006-2960

IS - 7

ER -

Cosper MM, Jameson GNL, Hernández HL, Krebs C, Huynh BH, Johnson MK. Characterization of the Cofactor Composition of Escherichia coli Biotin Synthase. Biochemistry. 2004 Feb 24;43(7):2007-2021. https://doi.org/10.1021/bi0356653