Physical and Functional Interactions of a Monothiol Glutaredoxin and an Iron Sulfur Cluster Carrier Protein with the Sulfur-donating Radical S-Adenosyl-l-methionine Enzyme MiaB

Boutigny, Sylvain; Saini, Avneesh; Baidoo, Edward E. K.; Yeung, Natasha; Keasling, Jay D.; Butland, Gareth

doi:10.1074/jbc.m113.460360

Cited by 28 publications

(22 citation statements)

References 36 publications

(19 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The NADH oxidase activity was not affected and the succinate oxidase activity was slightly reduced. GrxD is discussed as being part of repair machinery for damaged Fe/S clusters (Boutigny et al, 2013). Thus, it is most likely that GrxD acts as acceptor for damaged Fe/S clusters or as iron or sulfur donor for other chaperones.…”

Section: Discussionmentioning

confidence: 99%

Iron‐sulfur cluster carrier proteins involved in the assembly of Escherichia coli NADH:ubiquinone oxidoreductase (complex I)

Burschel

Decovic

Nuber

et al. 2018

Molecular Microbiology

View full text Add to dashboard Cite

Summary The NADH:ubiquinone oxidoreductase (respiratory complex I) is the main entry point for electrons into the Escherichia coli aerobic respiratory chain. With its sophisticated setup of 13 different subunits and 10 cofactors, it is anticipated that various chaperones are needed for its proper maturation. However, very little is known about the assembly of E. coli complex I, especially concerning the incorporation of the iron‐sulfur clusters. To identify iron‐sulfur cluster carrier proteins possibly involved in the process, we generated knockout strains of NfuA, BolA, YajL, Mrp, GrxD and IbaG that have been reported either to be involved in the maturation of mitochondrial complex I or to exert influence on the clusters of bacterial complex. We determined the NADH and succinate oxidase activities of membranes from the mutant strains to monitor the specificity of the individual mutations for complex I. The deletion of NfuA, BolA and Mrp led to a decreased stability and partially disturbed assembly of the complex as determined by sucrose gradient centrifugation and native PAGE. EPR spectroscopy of cytoplasmic membranes revealed that the BolA deletion results in the loss of the binuclear Fe/S cluster N1b.

show abstract

Section: Discussionmentioning

confidence: 99%

Iron‐sulfur cluster carrier proteins involved in the assembly of Escherichia coli NADH:ubiquinone oxidoreductase (complex I)

Burschel

Decovic

Nuber

et al. 2018

Molecular Microbiology

View full text Add to dashboard Cite

show abstract

“…These include so-called A-type carriers (ATC), 3 named IscA, SufA, and ErpA in prokaryotes and ISA1 and ISA2 in eukaryotes (13)(14)(15)(16)(17). Other carriers include the P-loop NTPases, (Ind1 in mitochondria, ApbC in Salmonella) (18 -21), the monothiol glutaredoxins (Grx 5 in yeast and GrxD in E. coli) (22)(23)(24), and the highly conserved NFU-type proteins that have been shown to interact with target proteins (25)(26)(27)(28)(29)(30)(31)(32)(33).…”

mentioning

confidence: 99%

The ErpA/NfuA complex builds an oxidation-resistant Fe-S cluster delivery pathway

Gerez

Huguenot

et al. 2018

Journal of Biological Chemistry

View full text Add to dashboard Cite

Fe-S cluster-containing proteins occur in most organisms, wherein they assist in myriad processes from metabolism to DNA repair via gene expression and bioenergetic processes. Here, we used both and methods to investigate the capacity of the four Fe-S carriers, NfuA, SufA, ErpA, and IscA, to fulfill their targeting role under oxidative stress. Likewise, Fe-S clusters exhibited varying half-lives, depending on the carriers they were bound to; an NfuA-bound Fe-S cluster was more stable ( = 100 min) than those bound to SufA ( = 55 min), ErpA ( = 54 min), or IscA ( = 45 min). Surprisingly, the presence of NfuA further enhanced stability of the ErpA-bound cluster to = 90 min. Using genetic and plasmon surface resonance analyses, we showed that NfuA and ErpA interacted directly with client proteins, whereas IscA or SufA did not. Moreover, NfuA and ErpA interacted with one another. Given all of these observations, we propose an architecture of the Fe-S delivery network in which ErpA is the last factor that delivers cluster directly to most if not all client proteins. NfuA is proposed to assist ErpA under severely unfavorable conditions. A comparison with the strategy employed in yeast and eukaryotes is discussed.

show abstract

“…Initial characterizations resulted in protein that had poor activity most likely due to incomplete cofactor incorporation which is essential for the enzyme’s activity. With regard to NfuA, the E. coli protein had been typically isolated aerobically and then subjected to a chemical reconstitution procedure to insert its iron-sulfur (Fe/S) cluster, leaving it unclear as to what the in vivo cluster configuration is (Angelini, et al, 2008; Py, et al, 2012; Boutigny, et al, 2013). Following the methods detailed herein, we have generated robust amounts of LipA and NfuA proteins that are >90% homogenous.…”

Section: Overproduction and Isolation Of E Coli Lipa And E Coli Nfuamentioning

confidence: 99%

Biochemical Approaches for Understanding Iron–Sulfur Cluster Regeneration in Escherichia coli Lipoyl Synthase During Catalysis

McCarthy

Booker

2018

Methods in Enzymology

View full text Add to dashboard Cite

Lipoyl synthase (LipA in bacteria) is a radical S-adenosylmethionine (SAM) enzyme that catalyzes the second step of the de novo biosynthesis of the lipoyl cofactor: the insertion of sulfur at C6 and C8 of a pendant octanoyl chain. In addition to the [4Fe4S] cluster that is characteristic of the radical SAM (RS) enzymes, LipA contains a second [4Fe4S] cluster that, though controversial, has been proposed to be degraded during turnover to supply the inserted sulfur atoms. A consequence of this proposed role is that the destruction of its iron-sulfur cluster renders the enzyme in an inactive state. Recently, it was shown that Escherichia coli proteins NfuA or IscU can confer catalytic properties to E. coli LipA in vitro. In this chapter, we present methods for characterizing LipA and analyzing its activity in vitro, and provide strategies to monitor the pathway for the regeneration of LipA's auxiliary cluster by E. coli iron-sulfur carrier protein NfuA.

show abstract

Physical and Functional Interactions of a Monothiol Glutaredoxin and an Iron Sulfur Cluster Carrier Protein with the Sulfur-donating Radical S-Adenosyl-l-methionine Enzyme MiaB

Cited by 28 publications

References 36 publications

Iron‐sulfur cluster carrier proteins involved in the assembly of Escherichia coli NADH:ubiquinone oxidoreductase (complex I)

Iron‐sulfur cluster carrier proteins involved in the assembly of Escherichia coli NADH:ubiquinone oxidoreductase (complex I)

The ErpA/NfuA complex builds an oxidation-resistant Fe-S cluster delivery pathway

Biochemical Approaches for Understanding Iron–Sulfur Cluster Regeneration in Escherichia coli Lipoyl Synthase During Catalysis

Contact Info

Product

Resources

About