Structural Insight into Concerted Inhibition of α2β2-Type Aspartate Kinase from Corynebacterium glutamicum

Yoshida, Ayako; Tanaka, Takeo; Kurihara, Takeshi; Fushinobu, Shinya; Kuzuyama, Tomohisa; Nishiyama, Makoto

doi:10.1016/j.jmb.2007.02.017

Cited by 43 publications

(81 citation statements)

References 38 publications

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“…Experiments using M. smegmatis showed that the deletion of the gene coding for AK results in significant cell growth inhibition (Rapaport et al, 1996). A negative regulatory feedback of AK by end products has been reported in Corynebacterium glutamicum (CgAK) and Thermus thermophilus (TtAK) (Cole et al, 1998;Yoshida et al, 2007a;Yoshida et al, 2009). Although no negative feedback regulation has been reported in MtbAK, a similar inhibitory regulation may exist in MtbAK.…”

Section: Introductionmentioning

confidence: 99%

“…AKI from Arabidopsis thaliana is an example of homo-oligomeric AK (Rognes et al, 1980;Mas-Droux et al, 2006;Schuldt et al, 2011). AKs from M. tuberculosis (Cirillo et al, 1994;Gilker J.M., 1997;Schuldt et al, 2011), C. glutamicum (Yoshida et al, 2007a) and T. thermophiles (Nishiyama et al, 1995;Yoshida et al, 2007b) belong to the second type. The hetero-tetrameric α 2 β 2 AK contains α and β subunits, which are encoded by in-frame overlapping genes (Schuldt et al, 2011).…”

Section: Introductionmentioning

confidence: 99%

“…The hetero-tetrameric α 2 β 2 AK contains α and β subunits, which are encoded by in-frame overlapping genes (Schuldt et al, 2011). The catalytic domain of heterotetrameric α 2 β 2 AK is composed of the N-terminal region of the α subunit and the regulatory domain comprises the β subunit and the C-terminal region of the α subunit (Gilker J.M., 1997;Yoshida et al, 2007aYoshida et al, , 2007bSchuldt et al, 2011). A key feature of the regulatory domain of AK is the conserved ACT domain [aspartate kinase, chorismate mutase, and TyrA (prephenate dehydrogenase)] (Aravind and Koonin, 1999;Chipman and Shaanan, 2001), which comprises a βαββαβ fold and is expected to play a regulatory role when binding to small molecules (Grant, 2006).…”

Section: Introductionmentioning

confidence: 99%

“…Crystal structures of several homo-oligomeric AKs and hetero-tetrameric α 2 β 2 -type AKs have been reported previously, such as the homo-oligomeric AK from A. thaliana (PDB code: 2CDQ) (Mas-Droux et al, 2006), the β subunit of α 2 β 2 -type AK from C. glutamicum (PDB code: 2DTJ) (Yoshida et al, 2007a), the β subunit of α 2 β 2 -type AK from T. thermophiles (TtAKβ) (PDB code: 2ZHO and 2DT9) (Yoshida et al, 2009) and the full-length α 2 β 2 -type AK from C. glutamicum (PDB code: 3AAW, 3AB2 and 3AB4) (Yoshida et al, 2010). However, no crystal structure of MtbAK has been reported and the regulatory mechanism of MtbAK is poorly understood.…”

Section: Introductionmentioning

confidence: 99%

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Structural view of the regulatory subunit of aspartate kinase from Mycobacterium tuberculosis

Yang

Yan

et al. 2011

Protein Cell

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The aspartate kinase (AK) from Mycobacterium tuberculosis (Mtb) catalyzes the biosynthesis of aspartate family amino acids, including lysine, threonine, isoleucine and methionine. We determined the crystal structures of the regulatory subunit of aspartate kinase from Mtb alone (referred to as MtbAKβ) and in complex with threonine (referred to as MtbAKβ-Thr) at resolutions of 2.6 Å and 2.0 Å, respectively. MtbAKβ is composed of two perpendicular non-equivalent ACT domains [aspartate kinase, chorismate mutase, and TyrA (prephenate dehydrogenase)] per monomer. Each ACT domain contains two α helices and four antiparallel β strands. The structure of MtbAKβ shares high similarity with the regulatory subunit of the aspartate kinase from Corynebacterium glutamicum (referred to as CgAKβ), suggesting similar regulatory mechanisms. Biochemical assays in our study showed that MtbAK is inhibited by threonine. Based on crystal structure analysis, we discuss the regulatory mechanism of MtbAK.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Structural view of the regulatory subunit of aspartate kinase from Mycobacterium tuberculosis

Yang

Yan

et al. 2011

Protein Cell

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“…A regulação das enzimas chave de vias multienzimáticas, chamadas de enzima reguladoras, ocorre através da ativação/inibição por seus produtos finais (YOSHIDA et al, 2010), processo chamado de retroalimentação positiva/retroalimentação negativa, que desempenha papel fundamental para a regulação da via e consequentemente na síntese dos seus aminoácidos produtos.…”

Section: Biossíntese De Lisina E Sua Regulaçãounclassified

Caracterização das enzimas chaves para o controle do metabolismo de lisina em milho (Zea mays L.) geneticamente modificado

Rizzi¹

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Analysis of intracellular metabolites of Corynebacterium glutamicum at high cell density with automated sampling and filtration and assessment of engineered enzymes for effective l‐lysine production

Luz

Hans

Frank

et al. 2016

Engineering in Life Sciences

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Engineering of enzymes and pathways is generally required for the development of efficient strains for bioproduction processes. To this end, quantitative and reliable data of intracellular metabolites are highly desired, but often not available, especially for conditions more close to industrial applications, i.e. at high cell density and product concentration. Here, we investigated the intracellular metabolite profiles of an engineered l‐lysine‐producing Corynebacterium glutamicum strain and the corresponding wild‐type strain to assess the impacts of deregulation of product inhibition of the key enzymes aspartate kinase and phosphoenolpyruvate carboxylase and to identify potentials for their further improvement. A bioreactor system with automated fast‐sampling, filtration and on‐filter quenching of the metabolism was used for a more reliable determination of intracellular metabolites in batch cultures with optical cell density (OD660) up to 40. The l‐lysine‐producing strain showed substantially different metabolite profiles in the amino acid metabolism, including increased intracellular pool sizes in the l‐lysine‐, l‐homoserine‐ and l‐threonine pathways and decreased intracellular pool sizes for all other determined amino acids. By comparing data of in vitro inhibition of the engineered enzymes and determined intracellular concentrations of the inhibitors it was found that the inferred in vivo activities of these enzymes are still significantly below their in vitro maximums. This work demonstrates the usefulness of metabolic analysis for assessing the impact of engineered enzymes and identifying targets for further strain development.

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Structural Insight into Concerted Inhibition of α2β2-Type Aspartate Kinase from Corynebacterium glutamicum

Cited by 43 publications

References 38 publications

Structural view of the regulatory subunit of aspartate kinase from Mycobacterium tuberculosis

Structural view of the regulatory subunit of aspartate kinase from Mycobacterium tuberculosis

Caracterização das enzimas chaves para o controle do metabolismo de lisina em milho (Zea mays L.) geneticamente modificado

Analysis of intracellular metabolites of Corynebacterium glutamicum at high cell density with automated sampling and filtration and assessment of engineered enzymes for effective l‐lysine production

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