NDLI: The iron-sulfur center of biotin synthase: site-directed mutants

Content Provider	Springer Nature Link
Author	Hewitson, Kirsty S. Ollagnier de Choudens, Sandrine Sanakis, Yiannis Shaw, Nicholas M. Baldwin, Jack E. Münck, Eckard Roach, Peter L. Fontecave, Marc
Copyright Year	2001
Abstract	Biotin synthase contains an essential [4Fe-4S]+ cluster that is thought to provide an electron for the cleavage of S-adenosylmethionine, a cofactor required for biotin formation. The conserved cysteine residues Cys53, Cys57 and Cys60 have been proposed as ligands to the [4Fe-4S] cluster. These residues belong to a C-X3-C-X2-C motif which is also found in pyruvate formate lyase-activating enzyme, lysine 2,3-aminomutase and the anaerobic ribonucleotide reductase-activating component. To investigate the role of the cysteine residues, Cys→Ala mutants of the eight cysteine residues of Escherichia coli biotin synthase were prepared and assayed for activity. Our results show that six cysteines are important for biotin formation. Only two mutant proteins, C276A and C288A, closely resembled the wild-type protein, indicating that the corresponding cysteines are not involved in iron chelation and biotin formation. The six other mutant proteins, C53A, C57A, C60A, C97A, C128A and C188A, were inactive but capable of assembling a [4Fe-4S] cluster, as shown by Mössbauer spectroscopy. The C53A, C57A and C60A mutant proteins are unique in that their cluster could not undergo reduction to the [4Fe-4S]+ state, as shown by EPR and Mössbauer spectroscopy. On this basis and by analogy with pyruvate formate lyase-activating enzyme and the anaerobic ribonucleotide reductase-activating component, it is suggested that the corresponding cysteines coordinate the cluster even though one cannot fully exclude the possibility that other cysteines play that role as well. Therefore it appears that for activity biotin synthase absolutely requires cysteines that are not involved in iron chelation.
Starting Page	83
Ending Page	93
Page Count	11
File Format	PDF
ISSN	09498257
Journal	JBIC Journal of Biological Inorganic Chemistry
Volume Number	7
Issue Number	1-2
e-ISSN	14321327
Language	English
Publisher	Springer Berlin Heidelberg
Publisher Date	2001-07-11
Publisher Place	Berlin, Heidelberg
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	Biochemistry Microbiology
Content Type	Text
Resource Type	Article
Subject	Biochemistry Inorganic Chemistry

Sl.	Authority	Responsibilities	Communication Details
1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
2	Indian Institute of Technology Kharagpur	Host Institute of the Project: The host institute of the project is responsible for providing infrastructure support and hosting the project	https://www.iitkgp.ac.in
3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
5	Website/Portal (Helpdesk)	Queries regarding NDLI and its services	support@ndl.gov.in
6	Contents and Copyright Issues	Queries related to content curation and copyright issues	content@ndl.gov.in
7	National Digital Library of India Club (NDLI Club)	Queries related to NDLI Club formation, support, user awareness program, seminar/symposium, collaboration, social media, promotion, and outreach	clubsupport@ndl.gov.in
8	Digital Preservation Centre (DPC)	Assistance with digitizing and archiving copyright-free printed books	dpc@ndl.gov.in
9	IDR Setup or Support	Queries related to establishment and support of Institutional Digital Repository (IDR) and IDR workshops	idr@ndl.gov.in

A substrate channel in the nitrogenase MoFe protein

Density functional theory calculations on the active site of biotin synthase: mechanism of S transfer from the Fe2S2 cluster and the role of 1st and 2nd sphere residues

Iron–sulfur protein folds, iron–sulfur chemistry, and evolution

Function of the tunnel in acetylcoenzyme A synthase/carbon monoxide dehydrogenase

Acetylene hydratase: a non-redox enzyme with tungsten and iron–sulfur centers at the active site

Structure-Function Studies of Escherichia coli Biotin Synthase via a Chemical Modification and Site-Directed Mutagenesis Approach

Iron–sulfur repair YtfE protein from Escherichia coli: structural characterization of the di-iron center

Identifying sequence determinants of reduction potentials of metalloproteins

Mechanisms of iron–sulfur cluster assembly: the SUF machinery

The iron-sulfur center of biotin synthase: site-directed mutants

Similar Documents

A substrate channel in the nitrogenase MoFe protein

Density functional theory calculations on the active site of biotin synthase: mechanism of S transfer from the Fe2S2 cluster and the role of 1st and 2nd sphere residues

Iron–sulfur protein folds, iron–sulfur chemistry, and evolution

Function of the tunnel in acetylcoenzyme A synthase/carbon monoxide dehydrogenase

Acetylene hydratase: a non-redox enzyme with tungsten and iron–sulfur centers at the active site

Structure-Function Studies of Escherichia coli Biotin Synthase via a Chemical Modification and Site-Directed Mutagenesis Approach

Iron–sulfur repair YtfE protein from Escherichia coli: structural characterization of the di-iron center

Identifying sequence determinants of reduction potentials of metalloproteins

Mechanisms of iron–sulfur cluster assembly: the SUF machinery

The iron-sulfur center of biotin synthase: site-directed mutants