Characterization of crystal structure and key residues of Aspergillus fumigatus nucleoside diphosphate kinase

Nucleoside selectivity of Aspergillus fumigatus nucleoside‐diphosphate kinase

“…In our work, we have solved crystal structures of unbound and NDP-bound (ADP, GDP, IDP, UDP, TDP and CDP) A. fumigatus NDK and assessed the oligomeric state of the enzyme using analytical-size exclusion chromatography (SEC) and native mass spectrometry. We present compelling evidence that A. fumigatus NDK forms a hexameric biological assembly, a finding which contradicts that of an earlier study [22]. It is vital to discern the true oligomeric state of A. fumigatus NDK amongst conflicting arguments as the quaternary structure is intimately tied to enzyme function.…”

“…In a previous paper, the crystal structure of A. fumigatus NDK was solved as a hexamer, arranged as a trimer of dimers [22]. However, analytical SEC indicated that the enzyme was tetrameric in solution [22]. Data obtained from the complementary techniques of crystallography, analytical SEC and native mass spectrometry unequivocally and consistently confirm that A. fumigatus NDK adopts a Table 3.…”

“…However, there is still an ongoing debate of the quaternary structure adopted by A. fumigatus NDK and how this relates to its biological function. In a previous paper, the crystal structure of A. fumigatus NDK was solved as a hexamer, arranged as a trimer of dimers [22]. However, analytical SEC indicated that the enzyme was tetrameric in solution [22].…”

“…Here we present the first study to elucidate the determinants of nucleoside selectivity in fungal NDK by performing a comprehensive analysis of the structure and enzymology of A. fumigatus NDK. The structure of A. fumigatus unbound‐NDK has been solved previously only in the unbound state, forming a crystallographic hexamer in the asymmetric unit but a tetramer in solution [22]. In our work, we have solved crystal structures of unbound and NDP‐bound (ADP, GDP, IDP, UDP, TDP and CDP) A. fumigatus NDK and assessed the oligomeric state of the enzyme using analytical‐size exclusion chromatography (SEC) and native mass spectrometry.…”

Identification, Characterization, and Site-Specific Mutagenesis of a Thermostable ω-Transaminase from Chloroflexi bacterium

“…Cb TA showed identity with amine transaminases from the following microorganisms: Arthrobacter sp. KNK168 (PDB ID: 3WWH; 38%), A. terreus NIH2624 (PDB ID: 4CE5; 36.1%) Aspergillus fumigatus Af293 (GenBank accession no. EAL86783; 37.2%), Archaeoglobus fulgidus DSM 4304 (PDB ID: 5MQZ; 37.8%), Nectria haematococca MPVI (PDB: 4CMD; 36.8%), M.…”