Domain Structure Characterization of the Multifunctional α-Aminoadipate Reductase from Penicillium chrysogenum by Limited Proteolysis

Hijarrubia, Maria Jose; Aparicio, Jesús F.; Martı́n, Juan F.

doi:10.1074/jbc.m211235200

Cited by 14 publications

(9 citation statements)

References 40 publications

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“…[22,23] The reaction is also reminiscent of that catalysed by aspartate semialdehyde dehydrogenase in bacteria, where aspartyl phosphate is the substrate for transthiolesterification and reduction. [24] Similar reductase domains in the myxochelin synthase catalyse two reductions to yield an alcohol.…”

Section: Resultsmentioning

confidence: 97%

Fusarin C Biosynthesis in Fusarium moniliforme and Fusarium venenatum

et al. 2004

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Fragments of polyketide synthase (PKS) genes were amplified from complementary DNA (cDNA) of the fusarin C producing filamentous fungi Fusarium moniliforme and Fusarium venenatum by using degenerate oligonucleotides designed to select for fungal PKS C-methyltransferase (CMeT) domains. The PCR products, which were highly homologous to fragments of known fungal PKS CMeT domains, were used to probe cDNA and genomic DNA (gDNA) libraries of F. moniliforme and F. venenatum. A 4.0 kb cDNA clone from F. venenatum was isolated and used to prepare a hygromycin-resistance knockout cassette, which was used to produce a fusarin-deficient strain of F. venenatum (kb = 1000 bp). Similarly, a 26 kb genomic fragment, isolated on two overlapping clones from F. moniliforme, encoded a complete iterative Type I PKS fused to an unusual nonribosomal peptide synthase module. Once again, targeted gene disruption produced a fusarin-deficient strain, thereby proving that this synthase is responsible for the first steps of fusarin biosynthesis.

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Section: Resultsmentioning

confidence: 97%

Fusarin C Biosynthesis in Fusarium moniliforme and Fusarium venenatum

et al. 2004

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“…Reminiscent of nonribosomal peptide synthetases (NRPSs), Lys2 reductases are organized as multidomain enzymes, each including an adenylation (A) domain, a thiolation (T) domain, which serves for covalent binding of the substrate, and a C‐terminal short‐chain reductase (R) domain (Figure 1). Notably, the A domain of a Lys2 enzyme is N‐terminally extended by an additional sequence of about 250 aa, the function of which has remained obscure 4. Hereafter, these domains are referred to as adenylation activating (ADA) domains.…”

Section: Methodsmentioning

confidence: 99%

Activity of α‐Aminoadipate Reductase Depends on the N‐Terminally Extending Domain

Kalb

Lackner²,

Rappe

et al. 2015

ChemBioChem

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L-α-Aminoadipic acid reductases catalyze the ATP- and NADPH-dependent reduction of L-α-aminoadipic acid to the corresponding 6-semialdehyde during fungal L-lysine biosynthesis. These reductases resemble peptide synthetases with regard to their multidomain composition but feature a unique domain of elusive function--now referred to as an adenylation activating (ADA) domain--that extends the reductase N-terminally. Truncated enzymes based on NPS3, the L-α-aminoadipic acid reductase of the basidiomycete Ceriporiopsis subvermispora, lacking the ADA domain either partially or entirely were tested for activity in vitro, together with an ADA-adenylation didomain and the ADA domainless adenylation domain. We provide evidence that the ADA domain is required for substrate adenylation: that is, the initial step of the catalytic turnover. Our biochemical data are supported by in silico modeling that identified the ADA domain as a partial peptide synthetase condensation domain.

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“…Primary protein sequences of A domains (for sequence accession numbers, see Table S1 in the supplemental material) were aligned separately using the MAFFT algorithm (18) implemented as a plugin for the Geneious software package version 7.0 (Biomatters). Sequences of both characterized (2,(8)(9)(10)(11) and uncharacterized reductases were chosen and retrieved from the NCBI protein database or by searching publically available genome sequences using BLAST (19). A rooted phylogenetic tree of A domains was inferred by Bayesian Inference using MrBayes 3.2 (20) using the following parameters: the mixed model of protein evolution was chosen as a prior (aamodelpr ϭ mixed).…”

Section: Methodsmentioning

confidence: 99%

“…1) which occurs with several peptide synthetases, coenzyme A (CoA) ligases, and other multifunctional proteins. Its significance has not been elucidated yet, but it may contribute to the structural integrity of the protein (9). Contrasting Lys2, other multidomain reductases lack this extra N-terminal extension and are therefore only about 1,000 to 1,050 aa in length.…”

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confidence: 99%

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Functional and Phylogenetic Divergence of Fungal Adenylate-Forming Reductases

Kalb

Lackner

Hoffmeister

2014

Appl Environ Microbiol

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A key step in fungal L-lysine biosynthesis is catalyzed by adenylate-forming L-␣-aminoadipic acid reductases, organized in domains for adenylation, thiolation, and the reduction step. However, the genomes of numerous ascomycetes and basidiomycetes contain an unexpectedly large number of additional genes encoding similar but functionally distinct enzymes. Here, we describe the functional in vitro characterization of four reductases which were heterologously produced in Escherichia coli. The Ceriporiopsis subvermispora serine reductase Nps1 features a terminal ferredoxin-NADP ؉ reductase (FNR) domain and thus belongs to a hitherto undescribed class of fungal multidomain enzymes. The second major class is characterized by the canonical terminal short-chain dehydrogenase/reductase domain and represented by Ceriporiopsis subvermispora Nps3 as the first biochemically characterized L-␣-aminoadipic acid reductase of basidiomycete origin. Aspergillus flavus L-tyrosine reductases LnaA and LnbA are members of a distinct phylogenetic clade. Phylogenetic analysis supports the view that fungal adenylate-forming reductases are more diverse than previously recognized and belong to four distinct classes.

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Domain Structure Characterization of the Multifunctional α-Aminoadipate Reductase from Penicillium chrysogenum by Limited Proteolysis

Cited by 14 publications

References 40 publications

Fusarin C Biosynthesis in Fusarium moniliforme and Fusarium venenatum

Fusarin C Biosynthesis in Fusarium moniliforme and Fusarium venenatum

Activity of α‐Aminoadipate Reductase Depends on the N‐Terminally Extending Domain

Functional and Phylogenetic Divergence of Fungal Adenylate-Forming Reductases

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