8PMS

NADase from Aspergillus fumigatus with replaced C-terminus from Neurospora crassa

8PMS の概要

• エントリーDOI: 10.2210/pdb8pms/pdb
• 関連するPDBエントリー: 6YGE 6YGF 6YGG 8PMR
• 分子名称: Conidial surface nicotinamide adenine dinucleotide glycohydrolase nadA, 2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-[alpha-L-fucopyranose-(1-6)]2-acetamido-2-deoxy-beta-D-glucopyranose, alpha-D-mannopyranose-(1-3)-beta-D-mannopyranose-(1-4)-2-acetamido-2-deoxy-beta-D-glucopyranose-(1-4)-[alpha-L-fucopyranose-(1-6)]2-acetamido-2-deoxy-beta-D-glucopyranose, ... (9 entities in total)
• 機能のキーワード: nadase, nad hydrolase, ca-binding, homodimer, glycoprotein, extracellular, hydrolase
• 由来する生物種: Aspergillus fumigatus Af293
• タンパク質・核酸の鎖数: 4
• 化学式量合計: 111526.37

構造登録者

Kallio, J.P.,Ferrario, E.,Stromland, O.,Ziegler, M. (登録日: 2023-06-29, 公開日: 2023-11-15, 最終更新日: 2024-11-06)

主引用文献

Ferrario, E.,Kallio, J.P.,Stromland, O.,Ziegler, M.
Novel Calcium-Binding Motif Stabilizes and Increases the Activity of Aspergillus fumigatus Ecto-NADase.
Biochemistry, 62:3293-3302, 2023

PubMed Abstract: Nicotinamide adenine dinucleotide (NAD) is an essential molecule in all kingdoms of life, mediating energy metabolism and cellular signaling. Recently, a new class of highly active fungal surface NADases was discovered. The enzyme from the opportunistic human pathogen was thoroughly characterized. It harbors a catalytic domain that resembles that of the tuberculosis necrotizing toxin from , which efficiently cleaves NAD to nicotinamide and ADP-ribose, thereby depleting the dinucleotide pool. Of note, the NADase has an additional Ca-binding motif at the C-terminus of the protein. Despite the presence of NADases in several fungal divisions, the Ca-binding motif is uniquely found in the Eurotiales order, which contains species that have immense health and economic impacts on humans. To identify the potential roles of the metal ion-binding site in catalysis or protein stability, we generated and characterized NADase variants lacking the ability to bind calcium. X-ray crystallographic analyses revealed that the mutation causes a drastic and dynamic structural rearrangement of the homodimer, resulting in decreased thermal stability. Even though the calcium-binding site is at a long distance from the catalytic center, the structural reorganization upon the loss of calcium binding allosterically alters the active site, thereby negatively affecting NAD-glycohydrolase activity. Together, these findings reveal that this unique calcium-binding site affects the protein fold, stabilizing the dimeric structure, but also mediates long-range effects resulting in an increased catalytic rate.

PubMed: 37934975
DOI: 10.1021/acs.biochem.3c00360

実験手法

X-RAY DIFFRACTION (2.4 Å)