Methionine Uptake in <i>Corynebacterium glutamicum</i> by MetQNI and by MetPS, a Novel Methionine and Alanine Importer of the NSS Neurotransmitter Transporter Family

Trötschel, Christian; Follmann, Martin; Nettekoven, Jeannine A.; Mohrbach, Tobias; Forrest, Lucy R.; Burkovski, Andreas; Marin, Kay; Krämer, Reinhard

doi:10.1021/bi801206t

Cited by 22 publications

(25 citation statements)

References 62 publications

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“…The genome of C. glutamicum carries approximately 750 membrane proteins that could possibly function as transporters, and members of 17 transporter families are predicted to be involved in amino acid transport (1). Among them, GluABCD, LysI, BrnQ, MetQNI, MetPS, PheP, and AroP have been functionally characterized (23)(24)(25)(26)(27)(28). AroP was identified as a general aromatic amino acid uptake system by gene disruption and complementation experiments (28).…”

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confidence: 99%

Characterization and Molecular Mechanism of AroP as an Aromatic Amino Acid and Histidine Transporter in Corynebacterium glutamicum

Shang

Zhang

et al. 2013

Journal of Bacteriology

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bCorynebacterium glutamicum is equipped with abundant membrane transporters to adapt to a changing environment. Many amino acid transporters have been identified in C. glutamicum, but histidine uptake has not been investigated in detail. Here, we identified the aromatic amino acid transporter encoded by aroP as a histidine transporter in C. glutamicum by a combination of the growth and histidine uptake features. Characterization of histidine uptake showed that AroP has a moderate affinity for histidine, with a K m value of 11.40 ؎ 2.03 M, and histidine uptake by AroP is competitively inhibited by the aromatic amino acids. Among the four substrates, AroP exhibits a stronger preference for tryptophan than for tyrosine, phenylalanine, and histidine. Homology structure modeling and molecular docking were performed to predict the substrate binding modes and conformational changes during substrate transport. These results suggested that tryptophan is best accommodated in the binding pocket due to shape compatibility, strong hydrophobic interactions, and the lowest binding energy, which is consistent with the observed substrate preference of AroP. Furthermore, the missense mutations of the putative substrate binding sites verified that Ser24, Ala28, and Gly29 play crucial roles in substrate binding and are highly conserved in the Gram-positive bacteria. Finally, the expression of aroP is not significantly affected by extracellular histidine or aromatic amino acids, indicating that the physiological role of AroP may be correlated with the increased fitness of C. glutamicum to assimilate extracellular amino acid for avoiding the high energy cost of amino acid biosynthesis.

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confidence: 99%

Characterization and Molecular Mechanism of AroP as an Aromatic Amino Acid and Histidine Transporter in Corynebacterium glutamicum

Shang

Zhang

et al. 2013

Journal of Bacteriology

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“…Methionine uptake transport and its regulation have been extensively studied in both E. coli and C. glutamicum (50,51). However, reports about methionine export are rare.…”

Section: Discussionmentioning

confidence: 99%

YjeH Is a Novel Exporter of l -Methionine and Branched-Chain Amino Acids in Escherichia coli

Liu

Liang

Zhang

et al. 2015

Appl Environ Microbiol

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c Amino acid efflux transport systems have important physiological functions and play vital roles in the fermentative production of amino acids. However, no methionine exporter has yet been identified in Escherichia coli. In this study, we identified a novel amino acid exporter, YjeH, in E. coli. The yjeH overexpression strain exhibited high tolerance to the structural analogues of Lmethionine and branched-chain amino acids, decreased intracellular amino acid levels, and enhanced export rates in the presence of a Met-Met, Leu-Leu, Ile-Ile, or Val-Val dipeptide, suggesting that YjeH functions as an exporter of L-methionine and the three branched-chain amino acids. The export of the four amino acids in the yjeH overexpression strain was competitively inhibited in relation to each other. The expression of yjeH was strongly induced by increasing cytoplasmic concentrations of substrate amino acids. Green fluorescent protein (GFP)-tagged YjeH was visualized by total internal reflection fluorescence microscopy to confirm the plasma membrane localization of YjeH. Phylogenetic analysis of transporters indicated that YjeH belongs to the amino acid efflux family of the amino acid/polyamine/organocation (APC) superfamily. Structural modeling revealed that YjeH has the typical "5 ؉ 5" transmembrane ␣-helical segment (TMS) inverted-repeat fold of APC superfamily transporters, and its binding sites are strictly conserved. The enhanced capacity of L-methionine export by the overexpression of yjeH in an L-methionine-producing strain resulted in a 70% improvement in titer. This study supplements the transporter classification and provides a substantial basis for the application of the methionine exporter in metabolic engineering. Membrane transport mediates the exchange of materials and energy with the surroundings to facilitate cell viability (1). Amino acids play central roles both as the building blocks of proteins and as intermediates in metabolism. Amino acid transport processes, including uptake and efflux, exist widely in bacteria. Amino acids in the surrounding environment can be imported into cells directly to participate in protein synthesis or carbon and nitrogen metabolism without spending energy for anabolism (2). The export process, however, is essential to maintain the intracellular amino acid pool and has significant applications for amino acid overproduction.Amino acid transport processes are mostly transporter mediated in prokaryotes. In Escherichia coli, the L-methionine uptake system MetNIQ is a typical primary transporter which utilizes the energy of ATP binding and hydrolysis to transport substrates (3), while the amino acid/polyamine/organocation (APC) superfamily is one of the largest superfamilies with nearly 250 secondary transporters. These transporters vary in length from 350 to 850 amino acid residues (4). Many amino acids are transported through APC carriers that function as solute-cation symporters and solute-solute antiporters, such as the lysine importer LysP in E. coli (5) and the aromatic amino ...

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“…BL1655 to BL1657 and BL1293 to BL1296 share similarities with MetNPQ, a high-affinity ABC transporter of B. subtilis (53,35, and 36% identities) (48) and a cystine ABC transporter of E. coli (48,31,33, and 26% identities), respectively. Other putative cystine or methionine transporters are present in the B. aurantiacum genome.…”

Section: Comparison Of the Expression Profiles After The Growth Of Stmentioning

confidence: 99%

“…The BL1249-BL1250-BL1251 proteins correspond to an ABC transporter that shares similarities with BL1293 to BL1296. BL3001 and BL3000 are 48 and 30% identical to MetPS, a low-affinity methionine transporter of C. glutamicum (53). We tested the effect of cysteine or methionine limitation on their expression.…”

Section: Comparison Of the Expression Profiles After The Growth Of Stmentioning

confidence: 99%

“…Moreover, ABC transporters and a symporter participate in the uptake of cystine in B. subtilis and Escherichia coli (9). Methionine can be taken up by highaffinity ABC transporters of the MUT family (48) or by a recently characterized MetPS symporter in Corynebacterium glutamicum (53). Finally, two different types of ABC transporters, the TauABCD and the SsuABC systems, are involved in sulfonate uptake (26,48,54).…”

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confidence: 99%

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Global Regulation of the Response to Sulfur Availability in the Cheese-Related Bacterium Brevibacterium aurantiacum

Forquin

Hébert

Roux

et al. 2011

Appl Environ Microbiol

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In this study, we combined metabolic reconstruction, growth assays, and metabolome and transcriptome analyses to obtain a global view of the sulfur metabolic network and of the response to sulfur availability in Brevibacterium aurantiacum. In agreement with the growth of B. aurantiacum in the presence of sulfate and cystine, the metabolic reconstruction showed the presence of a sulfate assimilation pathway, thiolation pathways that produce cysteine (cysE and cysK) or homocysteine (metX and metY) from sulfide, at least one gene of the transsulfuration pathway (aecD), and genes encoding three MetE-type methionine synthases. We also compared the expression profiles of B. aurantiacum ATCC 9175 during sulfur starvation or in the presence of sulfate. Under sulfur starvation, 690 genes, including 21 genes involved in sulfur metabolism and 29 genes encoding amino acids and peptide transporters, were differentially expressed. We also investigated changes in pools of sulfur-containing metabolites and in expression profiles after growth in the presence of sulfate, cystine, or methionine plus cystine. The expression of genes involved in sulfate assimilation and cysteine synthesis was repressed in the presence of cystine, whereas the expression of metX, metY, metE1, metE2, and BL613, encoding a probable cystathionine-␥-synthase, decreased in the presence of methionine. We identified three ABC transporters: two operons encoding transporters were transcribed more strongly during cysteine limitation, and one was transcribed more strongly during methionine depletion. Finally, the expression of genes encoding a methionine ␥-lyase (BL929) and a methionine transporter (metPS) was induced in the presence of methionine in conjunction with a significant increase in volatile sulfur compound production.

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Methionine Uptake in Corynebacterium glutamicum by MetQNI and by MetPS, a Novel Methionine and Alanine Importer of the NSS Neurotransmitter Transporter Family

Cited by 22 publications

References 62 publications

Characterization and Molecular Mechanism of AroP as an Aromatic Amino Acid and Histidine Transporter in Corynebacterium glutamicum

Characterization and Molecular Mechanism of AroP as an Aromatic Amino Acid and Histidine Transporter in Corynebacterium glutamicum

YjeH Is a Novel Exporter of l -Methionine and Branched-Chain Amino Acids in Escherichia coli

Global Regulation of the Response to Sulfur Availability in the Cheese-Related Bacterium Brevibacterium aurantiacum

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