Effect of iron–sulfur cluster assembly proteins on the expression of Escherichia coli lipoic acid synthase

Kriek, Marco; Peters, Liv; Takahashi, Yasahiro; Roach, Peter L.

doi:10.1016/s1046-5928(02)00680-0

Cited by 36 publications

(22 citation statements)

References 18 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Importantly, donation of sulfur from the [4Fe-4S] cluster within the enzyme suggests that LipA may be a self-sacrificing protein rather than performing true enzyme catalysis (5). This is supported by the small amount of LA generated per molecule of LipA in E. coli, which can be enhanced by coexpression of LipA with iron-sulfur cluster proteins (5,20). Recently, the E. coli iron-sulfur cluster proteins NfuA and IscU were shown to reinstall the [4Fe-4S] cluster of LipA in order to facilitate additional turns of the enzyme (21).…”

Section: Lipoic Acid Synthesismentioning

confidence: 99%

Lipoic acid metabolism and mitochondrial redox regulation

Solmonson

DeBerardinis

2018

Journal of Biological Chemistry

312

255

View full text Add to dashboard Cite

Lipoic acid is an essential cofactor for mitochondrial metabolism and is synthesized de novo using intermediates from mitochondrial fatty acid synthesis type II, S-adenosylmethionine and iron-sulfur clusters. This cofactor is required for catalysis by multiple mitochondrial 2-ketoacid dehydrogenase complexes, including pyruvate dehydrogenase, alpha-ketoglutarate dehydrogenase, and branched-chain ketoacid dehydrogenase. Lipoic acid also plays a critical role in stabilizing and regulating these multienzyme complexes. Many of these dehydrogenases are regulated by reactive oxygen species, mediated through the disulfide bond of the prosthetic lipoyl moiety.Collectively, its functions explain why lipoic acid is required for cell growth, mitochondrial activity and coordination of fuel metabolism.

show abstract

Section: Lipoic Acid Synthesismentioning

confidence: 99%

Lipoic acid metabolism and mitochondrial redox regulation

Solmonson

DeBerardinis

2018

Journal of Biological Chemistry

312

255

View full text Add to dashboard Cite

show abstract

“…If results analogous to those of BioB are obtained, the question of how the novel LipA CXXXXCXXXXXC motif becomes liganded by a [4Fe-4S] cluster. In this regard coexpression of LipA with the Isc proteins results in increased LipA activity (254). However, it remains to be seen which clusters are built by this manipulation.…”

Section: Remaining Questions In Lipoic Acid Synthesismentioning

confidence: 99%

Biotin and Lipoic Acid: Synthesis, Attachment, and Regulation

Cronan

2014

EcoSal Plus

View full text Add to dashboard Cite

Summary Two vitamins, biotin and lipoic acid, are essential in all three domains of life. Both coenzymes function only when covalently attached to key metabolic enzymes. There they act as “swinging arms” that shuttle intermediates between two active sites (= covalent substrate channeling) of key metabolic enzymes. Although biotin was discovered over 100 years ago and lipoic acid 60 years ago, it was not known how either coenzyme is made until recently. In Escherichia coli the synthetic pathways for both coenzymes have now been worked out for the first time. The late steps of biotin synthesis, those involved in assembling the fused rings, were well-described biochemically years ago, although recent progress has been made on the BioB reaction, the last step of the pathway in which the biotin sulfur moiety is inserted. In contrast, the early steps of biotin synthesis, assembly of the fatty acid-like “arm” of biotin were unknown. It has now been demonstrated that the arm is made by using disguised substrates to gain entry into the fatty acid synthesis pathway followed by removal of the disguise when the proper chain length is attained. The BioC methyltransferase is responsible for introducing the disguise and the BioH esterase for its removal. In contrast to biotin, which is attached to its cognate proteins as a finished molecule, lipoic acid is assembled on its cognate proteins. An octanoyl moiety is transferred from the octanoyl-ACP of fatty acid synthesis to a specific lysine residue of a cognate protein by the LipB octanoyl transferase followed by sulfur insertion at carbons C6 and C8 by the LipA lipoyl synthetase. Assembly on the cognate proteins regulates the amount of lipoic acid synthesized and thus there is no transcriptional control of the synthetic genes. In contrast transcriptional control of the biotin synthetic genes is wielded by a remarkably sophisticated, yet simple, system, exerted through BirA a dual function protein that both represses biotin operon transcription and ligates biotin to its cognate protein.

show abstract

“…The K144A variant was generated from the wild-type MtAPR template using a QuikChange sitedirected mutagenesis kit (Stratagene, La Jolla, CA) and the following primer sequence: 5Ј-GCTGCCGGTTGCGCAAG-GTCGTTCCCCTGGG-3Ј. Plasmids encoding wild-type, C256S, or K144A MtAPR pET24 and pACYC (containing genes encoding the isc operon of six accessory proteins required for iron-sulfur cluster biosynthesis in Azotobacter vinelandii under the control of an arabinose-inducible promoter) (25) were cotransformed into Escherichia coli BL21(DE3) (Novagen, San Diego, CA) and plated on L-agar 50 g/ml kanamycin and 100 g/ml carbenicillin. A single colony was picked and added to 5 ml of L-broth plus antibiotics and grown overnight with shaking at 37°C.…”

Section: Methodsmentioning

confidence: 99%

“…However, yields from these preparations were low because of the large quantity of MtAPR present in the insoluble protein fraction. To improve the yield and stability of purified MtAPR, we coexpressed the MtAPR gene with the gene products of the isc operon required for iron-sulfur cluster biosynthesis in A. vinelandii (25). Under these conditions the yield of MtAPR was typically 7 mg/liter of culture, which represents an improvement over the ϳ1 mg/liter obtained when MtAPR is overexpressed in the absence of the isc proteins.…”

Section: Purification and Spectroscopic Characterization Of The [4fe-4s]mentioning

confidence: 99%

Spectroscopic Studies on the [4Fe-4S] Cluster in Adenosine 5′-Phosphosulfate Reductase from Mycobacterium tuberculosis

Bhave¹,

Hong

Lee³

et al. 2011

Journal of Biological Chemistry

View full text Add to dashboard Cite

Mycobacterium tuberculosis adenosine 5-phosphosulfate reductase (MtAPR) is an iron-sulfur protein and a validated target to develop new antitubercular agents, particularly for the treatment of latent infection. The enzyme harbors a [4Fe-4S]2؉ cluster that is coordinated by four cysteinyl ligands, two of which are adjacent in the amino acid sequence. The ironsulfur cluster is essential for catalysis; however, the precise role of the [4Fe-4S] cluster in APR remains unknown. Progress in this area has been hampered by the failure to generate a paramagnetic state of the [4Fe-4S] cluster that can be studied by electron paramagnetic resonance spectroscopy. Herein, we overcome this limitation and report the EPR spectra of MtAPR in the [4Fe-4S] ؉ state. The EPR signal is rhombic and consists of two overlapping S ‫؍‬ 1 ⁄ 2 species. Substrate binding to MtAPR led to a marked increase in the intensity and resolution of the EPR signal and to minor shifts in principle g values that were not observed among a panel of substrate analogs, including adenosine 5-diphosphate. Using site-directed mutagenesis, in conjunction with kinetic and EPR studies, we have also identified an essential role for the active site residue Lys-144, whose side chain interacts with both the iron-sulfur cluster and the sulfate group of adenosine 5-phosphosulfate. The implications of these findings are discussed with respect to the role of the iron-sulfur cluster in the catalytic mechanism of APR.In bacteria and plants, activation of inorganic sulfur is required for de novo biosynthesis of cysteine. To this end, the metabolic assimilation of sulfate from the environment proceeds via adenosine 5Ј-phosphosulfate (APS) 2 or 3Ј-phosphoadenosine-5Ј-phosphosulfate (PAPS) (1). These intermediates are produced by the action of ATP-sulfurylase (EC 2.7.7.4), which condenses sulfate and ATP to form APS (2, 3), and by APS kinase (EC 2.7.1.25), which produces PAPS from ATP and APS (4).APS and PAPS are reduced by enzymes in the reductive branch of the sulfate assimilation pathway, producing sulfite and AMP or adenosine 3Ј,5Ј-diphosphate (Scheme 1). These enzymes can be subdivided into two groups according to their substrate preference: the APS reductases (APR) and the PAPS reductases (PAPR) (EC 1.8.99.4). Functional and structural studies have been used to investigate the chemical reaction mechanism of APR and PAPR enzymes (1,(5)(6)(7)(8). The mechanism involves nucleophilic attack by the active site cysteine on the sulfur atom of APS or PAPS to form an enzyme S-sulfocysteine intermediate, which is cleaved by thiol-disulfide exchange with thioredoxin or glutaredoxin (Fig. 1). The sulfite product is then reduced to sulfide by sulfite reductase (EC 1.8.7.1) and utilized to synthesize cysteine and other essential sulfur-containing biomolecules (9). In the human pathogen Mycobacterium tuberculosis, APR is a validated target against the latent phase of infection (10).Only a 3Ј-phosphate group distinguishes PAPS from APS. Accordingly, APR and PAPR have nearly identical three...

show abstract

Effect of iron–sulfur cluster assembly proteins on the expression of Escherichia coli lipoic acid synthase

Cited by 36 publications

References 18 publications

Lipoic acid metabolism and mitochondrial redox regulation

Lipoic acid metabolism and mitochondrial redox regulation

Biotin and Lipoic Acid: Synthesis, Attachment, and Regulation

Spectroscopic Studies on the [4Fe-4S] Cluster in Adenosine 5′-Phosphosulfate Reductase from Mycobacterium tuberculosis

Contact Info

Product

Resources

About