Novel Posttranslational Activation of the
            <i>LYS2-</i>
            Encoded α-Aminoadipate Reductase for Biosynthesis of Lysine and Site-Directed Mutational Analysis of Conserved Amino Acid Residues in the Activation Domain of
            <i>Candida albicans</i>

Guo, Shujuan; Evans, Sarah; Wilkes, Mindy B.; Bhattacharjee, J. K.

doi:10.1128/jb.183.24.7120-7125.2001

Cited by 22 publications

(31 citation statements)

References 35 publications

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“…Their specific feature is a 4Ј-PP component covalently attached to a conserved serine residue through a phosphodiester bond (24). Another enzyme with a 4Ј-PP modification is ␣-aminoadipate reductase, a component of lysine synthesis specific to fungi (32). The 4Ј-PP arm is a crucial functional component of the carrier proteins: it holds a growing chain of fatty acids, polyketides, or peptides during the reaction.…”

Section: Discussionmentioning

confidence: 99%

10-Formyltetrahydrofolate Dehydrogenase Requires a 4′-Phosphopantetheine Prosthetic Group for Catalysis

Donato

Krupenko

Tsybovsky

et al. 2007

Journal of Biological Chemistry

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10-Formyltetrahydrofolate dehydrogenase (FDH) consists of two independent catalytic domains, N-and C-terminal, connected by a 100-amino acid residue linker (intermediate domain). Our previous studies on structural organization and enzymatic properties of rat FDH suggest that the overall enzyme reaction, i.e. NADP ؉ -dependent conversion of 10-formyltetrahydrofolate to tetrahydrofolate and CO 2 , consists of two steps: (i) hydrolytic cleavage of the formyl group in the N-terminal catalytic domain, followed by (ii) NADP ؉ -dependent oxidation of the formyl group to CO 2 in the C-terminal aldehyde dehydrogenase domain. In this mechanism, it was not clear how the formyl group is transferred between the two catalytic domains after the first step. This study demonstrates that the intermediate domain functions similarly to an acyl carrier protein. The subunit of 10-formyltetrahydrofolate dehydrogenase (FDH 3 ; ALDH1L1; EC 1.5.1.6) represents a single polypeptide consisting of three domains, with two of these domains possessing their own catalytic activities (1-4). The N-terminal domain of FDH (residues 1-310) functions as 10-formyltetrahydrofolate (10-fTHF) hydrolase, converting 10-fTHF to tetrahydrofolate (THF) and formate ( Fig. 1) (2). The C-terminal domain (residues 420 -902) is an aldehyde dehydrogenase-homologous enzyme capable of NADP ϩ -dependent oxidation of short chain aldehydes to the corresponding acids (3). The two catalytic domains are separated by a short (ϳ100 residues) linker domain (intermediate domain). Although the physiological significance of the catalytic functions of the separate domains of FDH is not clear, the well established function of FDH is the conversion of 10-fTHF to THF in an NADP ϩ -dependent dehydrogenase reaction (5-7). It is believed that this reaction regulates intracellular 10-fTHF/THF pools (8), controls de novo purine biosynthesis (9, 10), and affects the methylation potential of the cell (11). This reaction is observed only when the two catalytic domains are linked in one polypeptide by the intermediate domain (2). Studies of FDH structure and catalytic mechanism (12-18) suggest that this reaction proceeds in two steps: (i) hydrolytic removal of the formyl group from 10-fTHF and (ii) NADP ϩ -dependent oxidation of formyl to CO 2 . The first step takes place in the N-terminal hydrolase domain, whereas the second step occurs in the C-terminal aldehyde dehydrogenase domain (2,3,17,18).The connection between the two steps is the transfer of the formyl group between the two catalytic domains. However, the nature of such a transfer is not obvious. We have demonstrated previously that the two catalytic domains of FDH do not form close contacts in the absence of the intermediate domain (2). It has been suggested that the intermediate domain keeps the N-and C-terminal domains of FDH in close proximity to each other and in the correct orientation to create an interface between the two domains and to allow the formyl group transfer between the two catalytic centers. In support of this, t...

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Section: Discussionmentioning

confidence: 99%

10-Formyltetrahydrofolate Dehydrogenase Requires a 4′-Phosphopantetheine Prosthetic Group for Catalysis

Donato

Krupenko

Tsybovsky

et al. 2007

Journal of Biological Chemistry

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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%

“…It is the first characterized basidiomycete representative and, along with, e.g., Lys2 of S. cerevisiae and Ca. albicans (8,31,33), is one out of only very few examples that have been characterized biochemically.…”

Section: L-tyrosine Reductase A-t-sdr (Class Ii) L-α-aminoadipic Acidmentioning

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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“…This intermediate is then converted into ␣-AA-␦-semialdehyde by the action of the ␣-aminoadipate reductase (␣-AAR, EC 1.2.1.31) encoded by the lys2 and lys5 genes. The product of the lys2 gene constitutes the apoenzyme, whereas the lys5 product appears to be a specific phosphopantetheinyl transferase for post-translational modification of Lys2 (5,6). The ␣-AAR, also called ␣-aminoadipate semialdehyde dehydrogenase, first activates the ␣-AA ␦-carboxyl group by an ATPdependent process through the formation of an ␣-AA-adenylate, a unique step among amino acid biosynthetic pathways, that is then reduced by the reduction domain using NADPH to yield ␣-AA-␦-semialdehyde and AMP.…”

mentioning

confidence: 99%

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

Hijarrubia

Aparicio

Martı́n

2003

Journal of Biological Chemistry

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The ␣-aminoadipate reductase (␣-AAR) of Penicillium chrysogenum, an enzyme that activates the ␣-aminoadipic acid by forming an ␣-aminoadipyl adenylate and reduces the activated intermediate to ␣-aminoadipic semialdehyde, was purified to homogeneity by immunoaffinity techniques, and the kinetics for ␣-aminoadipic acid, ATP, and NADPH were determined. Sequencing of the N-terminal end confirmed the 10 first amino acids deduced from the nucleotide sequence. Its domain structure has been investigated using limited proteolysis and active site labeling. Trypsin and elastase were used to cleave the multienzyme, and the location of fragments within the

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Novel Posttranslational Activation of the LYS2- Encoded α-Aminoadipate Reductase for Biosynthesis of Lysine and Site-Directed Mutational Analysis of Conserved Amino Acid Residues in the Activation Domain of Candida albicans

Cited by 22 publications

References 35 publications

10-Formyltetrahydrofolate Dehydrogenase Requires a 4′-Phosphopantetheine Prosthetic Group for Catalysis

10-Formyltetrahydrofolate Dehydrogenase Requires a 4′-Phosphopantetheine Prosthetic Group for Catalysis

Functional and Phylogenetic Divergence of Fungal Adenylate-Forming Reductases

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

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