5LS0

Crystal structure of Inorganic Pyrophosphatase PPA1 from Arabidopsis thaliana

5LS0 の概要

• エントリーDOI: 10.2210/pdb5ls0/pdb
• 関連するPDBエントリー: 4lug
• 分子名称: Soluble inorganic pyrophosphatase 1, MAGNESIUM ION, DI(HYDROXYETHYL)ETHER, ... (4 entities in total)
• 機能のキーワード: inorganic pyrophosphatase, hydrolase, ob-fold
• 由来する生物種: Arabidopsis thaliana (Mouse-ear cress)
• タンパク質・核酸の鎖数: 2
• 化学式量合計: 41299.82

構造登録者

Grzechowiak, M.,Sikorski, M.,Jaskolski, M. (登録日: 2016-08-22, 公開日: 2017-09-13, 最終更新日: 2024-01-17)

主引用文献

Grzechowiak, M.,Ruszkowski, M.,Sliwiak, J.,Szpotkowski, K.,Sikorski, M.,Jaskolski, M.
Crystal structures of plant inorganic pyrophosphatase, an enzyme with a moonlighting autoproteolytic activity.
Biochem.J., 476:2297-2319, 2019

PubMed Abstract: Inorganic pyrophosphatases (PPases, EC 3.6.1.1), which hydrolyze inorganic pyrophosphate to phosphate in the presence of divalent metal cations, play a key role in maintaining phosphorus homeostasis in cells. DNA coding inorganic pyrophosphatases from (PPA1) and (PPA1) were cloned into a bacterial expression vector and the proteins were produced in cells and crystallized. In terms of their subunit fold, PPA1 and PPA1 are reminiscent of other members of Family I soluble pyrophosphatases from bacteria and yeast. Like their bacterial orthologs, both plant PPases form hexamers, as confirmed in solution by multi-angle light scattering and size-exclusion chromatography. This is in contrast with the fungal counterparts, which are dimeric. Unexpectedly, the crystallized PPA1 and PPA1 proteins lack ∼30 amino acid residues at their N-termini, as independently confirmed by chemical sequencing. , self-cleavage of the recombinant proteins is observed after prolonged storage or during crystallization. The cleaved fragment corresponds to a putative signal peptide of mitochondrial targeting, with a predicted cleavage site at Val31-Ala32. Site-directed mutagenesis shows that mutations of the key active site Asp residues dramatically reduce the cleavage rate, which suggests a moonlighting proteolytic activity. Moreover, the discovery of autoproteolytic cleavage of a mitochondrial targeting peptide would change our perception of this signaling process.

PubMed: 31371393
DOI: 10.1042/BCJ20190427

実験手法

X-RAY DIFFRACTION (1.83 Å)