5DAZ

Crystal structure of Scabin, a mono-ADP-ribosyltransferase from Streptomyces scabies

5DAZ の概要

• エントリーDOI: 10.2210/pdb5daz/pdb
• 分子名称: Scabin (2 entities in total)
• 機能のキーワード: mono-adp-ribosyltransferase, apo-enzyme, nad-binding protein, transferase
• 由来する生物種: Streptomyces scabiei 87.22
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 21857.17

構造登録者

Dutta, D.,Lanoue, J.,Merrill, A.R. (登録日: 2015-08-20, 公開日: 2016-03-23, 最終更新日: 2024-10-09)

主引用文献

Lyons, B.,Ravulapalli, R.,Lanoue, J.,Lugo, M.R.,Dutta, D.,Carlin, S.,Merrill, A.R.
Scabin, a Novel DNA-acting ADP-ribosyltransferase from Streptomyces scabies.
J.Biol.Chem., 291:11198-11215, 2016

PubMed Abstract: A bioinformatics strategy was used to identify Scabin, a novel DNA-targeting enzyme from the plant pathogen 87.22 strain of Streptomyces scabies Scabin shares nearly 40% sequence identity with the Pierisin family of mono-ADP-ribosyltransferase toxins. Scabin was purified to homogeneity as a 22-kDa single-domain enzyme and was shown to possess high NAD(+)-glycohydrolase (Km (NAD) = 68 ± 3 μm; kcat = 94 ± 2 min(-1)) activity with an RSQXE motif; it was also shown to target deoxyguanosine and showed sigmoidal enzyme kinetics (K0.5(deoxyguanosine) = 302 ± 12 μm; kcat = 14 min(-1)). Mass spectrometry analysis revealed that Scabin labels the exocyclic amino group on guanine bases in either single-stranded or double-stranded DNA. Several small molecule inhibitors were identified, and the most potent compounds were found to inhibit the enzyme activity with Ki values ranging from 3 to 24 μm PJ34, a well known inhibitor of poly-ADP-ribosyltransferases, was shown to be the most potent inhibitor of Scabin. Scabin was crystallized, representing the first structure of a DNA-targeting mono-ADP-ribosyltransferase enzyme; the structures of the apo-form (1.45 Å) and with two inhibitors (P6-E, 1.4 Å; PJ34, 1.6 Å) were solved. These x-ray structures are also the first high resolution structures of the Pierisin subgroup of the mono-ADP-ribosyltransferase toxin family. A model of Scabin with its DNA substrate is also proposed.

PubMed: 27002155
DOI: 10.1074/jbc.M115.707653

実験手法

X-RAY DIFFRACTION (1.45 Å)