TY  - JOUR
A1  - Tiedemann, Kim
A1  - Iobbi-Nivol, Chantal
A1  - Leimkühler, Silke
T1  - The Role of the Nucleotides in the Insertion of the bis-Molybdopterin Guanine Dinucleotide Cofactor into apo-Molybdoenzymes
JF  - Molecules
N2  - The role of the GMP nucleotides of the bis-molybdopterin guanine dinucleotide (bis-MGD) cofactor of the DMSO reductase family has long been a subject of discussion. The recent characterization of the bis-molybdopterin (bis-Mo-MPT) cofactor present in the E. coli YdhV protein, which differs from bis-MGD solely by the absence of the nucleotides, now enables studying the role of the nucleotides of bis-MGD and bis-MPT cofactors in Moco insertion and the activity of molybdoenzymes in direct comparison. Using the well-known E. coli TMAO reductase TorA as a model enzyme for cofactor insertion, we were able to show that the GMP nucleotides of bis-MGD are crucial for the insertion of the bis-MGD cofactor into apo-TorA.
KW  - bis-MGD
KW  - chaperone
KW  - molybdenum cofactor
KW  - TMAO reductase
Y1  - 2022
U6  - https://doi.org/10.3390/molecules27092993
SN  - 1420-3049
VL  - 27
SP  - 1
EP  - 15
PB  - MDPI
CY  - Basel, Schweiz
ET  - 9
ER  - 
TY  - JOUR
A1  - Ogunkola, Moses Olalekan
A1  - Guiraudie-Capraz, Gaelle
A1  - Féron, François
A1  - Leimkühler, Silke
T1  - The Human Mercaptopyruvate Sulfurtransferase TUM1 Is Involved in Moco Biosynthesis, Cytosolic tRNA Thiolation and Cellular Bioenergetics in Human Embryonic Kidney Cells
JF  - Biomolecules
N2  - Sulfur is an important element that is incorporated into many biomolecules in humans. The incorporation and transfer of sulfur into biomolecules is, however, facilitated by a series of different sulfurtransferases. Among these sulfurtransferases is the human mercaptopyruvate sulfurtransferase (MPST) also designated as tRNA thiouridine modification protein (TUM1). The role of the human TUM1 protein has been suggested in a wide range of physiological processes in the cell among which are but not limited to involvement in Molybdenum cofactor (Moco) biosynthesis, cytosolic tRNA thiolation and generation of H2S as signaling molecule both in mitochondria and the cytosol. Previous interaction studies showed that TUM1 interacts with the L-cysteine desulfurase NFS1 and the Molybdenum cofactor biosynthesis protein 3 (MOCS3). Here, we show the roles of TUM1 in human cells using CRISPR/Cas9 genetically modified Human Embryonic Kidney cells. Here, we show that TUM1 is involved in the sulfur transfer for Molybdenum cofactor synthesis and tRNA thiomodification by spectrophotometric measurement of the activity of sulfite oxidase and liquid chromatography quantification of the level of sulfur-modified tRNA. Further, we show that TUM1 has a role in hydrogen sulfide production and cellular bioenergetics.
KW  - Moco biosynthesis
KW  - sulfite oxidase
KW  - cytosolic tRNA thiolation
KW  - 5-methoxycarbonylmethyl-2-thiouridine
KW  - H2S biosynthesis
KW  - cellular bioenergetics
Y1  - 2023
U6  - https://doi.org/10.3390/biom13010144
SN  - 2218-273X
VL  - 13
SP  - 1
EP  - 23
PB  - MDPI
CY  - Basel, Schweiz
ET  - 1
ER  -