Effect of lysine succinylation on the regulation of 2-oxoglutarate dehydrogenase inhibitor, OdhI, involved in glutamate production inCorynebacterium glutamicum

Abstract

In Corynebacterium glutamicum, the activity of the 2-oxoglutarate dehydrogenase (ODH) complex is negatively regulated by the unphosphorylated form of OdhI protein, which is critical for L-glutamate overproduction. We examined the potential impact of protein acylation at lysine (K)-132 of OdhI in C. glutamicum ATCC13032. The K132E succinylation-mimic mutation reduced the ability of OdhI to bind OdhA, the catalytic subunit of the ODH complex, which reduced the inhibition of ODH activity. In vitro succinylation of OdhI protein also reduced the ability to inhibit ODH, and the K132R mutation blocked the effect. These results suggest that succinylation at K132 may attenuate the OdhI function. Consistent with these results, the C. glutamicum mutant strain with OdhI-K132E showed decreased L-glutamate production. Our results indicated that not only phosphorylation but also succinylation of OdhI protein may regulate L-glutamate production in C. glutamicum.

Elevated lysine crotonylation and succinylation in the brains of BTBR mice

“…Furthermore, the evoked glutamate release in the cerebral cortex of BTBR mice was certified to be distinctly lower than that in B6 mice (Wei et al, 2016a). Succinylation is closely associated with glutamate production, and the study supports the notion that succinylation inhibits glutamate production in corynebacterium glutamicum (Komine‐Abe et al, 2017). The increased lysine succinylation of proteins may partly explain the deficit in glutamate release in BTBR mice.…”

In vitro reconstitution and characterization of pyruvate dehydrogenase and 2‐oxoglutarate dehydrogenase hybrid complex from Corynebacterium glutamicum

“…Recombinant Cg E1o protein was expressed as a C‐terminal His‐tagged protein using the pET‐21a vector (pAA62) (Komine‐Abe et al, 2017). Recombinant Cg E1p protein was expressed as a C‐terminal His‐ and Flag‐tagged protein using the pET‐21a vector (pKH4).…”

“…N‐terminal His‐ and Flag‐tagged Cg E3 enabled copurification of Cg E2 and Cg E3 as a Cg E2‐E3 subcomplex. Recombinant OdhI protein was expressed as an N‐terminal Strep‐tagged protein (pAA77) (Komine‐Abe et al, 2017).…”

“…Pyruvate dehydrogenase (PDH) and 2‐oxoglutarate dehydrogenase (ODH) are critical enzymes in central carbon metabolism and generate vital high energy compounds acetyl‐CoA and succinyl‐CoA, respectively; both of which are major CoA derivatives in cell and serve as substrates for protein acetylation and succinylation (Komine‐Abe et al, 2017; Mizuno et al, 2016; Nagano‐Shoji et al, 2017). PDH and ODH are large enzyme complexes (∼10 MDa) composed of three subunits; E1, E2, and E3.…”

Large-Scale Assessment of Bioinformatics Tools for Lysine Succinylation Sites

“…The identification of protein succinylation sites is a crucial topic in cellular pathology and physiology, which may provide valuable information for biomedical research and drug development. In recent years, high-throughput methods with mass spectrometry and succinylation enrichment analysis have been extensively implemented to identify lysine succinylation in several organisms [1,2,7,22,25,37,45,46,47,48,49]. A large-scale protein lysine-succinylated sites have been verified by experimentally in both prokaryotes [7,24,50,51] and eukaryotes [2,24,25,47].…”