2O0V

Pectate lyase bound to hexasaccharide compound III

2O0V の概要

• エントリーDOI: 10.2210/pdb2o0v/pdb
• 関連するPDBエントリー: 1BN8 2NZM 2O04 2O0W 2O17 2O1D
• 分子名称: Pectate lyase, alpha-D-galactopyranuronic acid-(1-4)-alpha-D-galactopyranuronic acid-(1-4)-alpha-D-galactopyranuronic acid-(1-4)-alpha-D-galactopyranuronic acid-(1-4)-alpha-D-galactopyranuronic acid, CALCIUM ION, ... (4 entities in total)
• 機能のキーワード: michaelis complex with compound iii, lyase
• 由来する生物種: Bacillus subtilis
• 細胞内の位置: Secreted: P39116
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 44377.94

構造登録者

Pickersgill, R.W.,Seyedarabi, A. (登録日: 2006-11-28, 公開日: 2007-11-20, 最終更新日: 2023-12-27)

主引用文献

Seyedarabi, A.,To, T.T.,Ali, S.,Hussain, S.,Fries, M.,Madsen, R.,Clausen, M.H.,Teixteira, S.,Brocklehurst, K.,Pickersgill, R.W.
Structural insights into substrate specificity and the anti beta-elimination mechanism of pectate lyase.
Biochemistry, 49:539-546, 2010

PubMed Abstract: Pectate lyases harness anti beta-elimination chemistry to cleave the alpha-1,4 linkage in the homogalacturonan region of plant cell wall pectin. We have studied the binding of five pectic oligosaccharides to Bacillus subtilis pectate lyase in crystals of the inactive enzyme in which the catalytic base is substituted with alanine (R279A). We discover that the three central subsites (-1, +1, and +2) have a profound preference for galacturonate but that the distal subsites can accommodate methylated galacturonate. It is reasonable to assume therefore that pectate lyase can cleave pectin with three consecutive galacturonate residues. The enzyme in the absence of substrate binds a single calcium ion, and we show that two additional calcium ions bind between enzyme and substrate carboxylates occupying the +1 subsite in the Michaelis complex. The substrate binds less intimately to the enzyme in a complex made with a catalytic base in place but in the absence of the calcium ions and an adjacent lysine. In this complex, the catalytic base is correctly positioned to abstract the C5 proton, but there are no calcium ions binding the carboxylate at the +1 subsite. It is clear, therefore, that the catalytic calcium ions and adjacent lysine promote catalysis by acidifying the alpha-proton, facilitating its abstraction by the base. There is also clear evidence that binding distorts the relaxed 2(1) or 3(1) helical conformation of the oligosaccharides in the region of the scissile bond.

PubMed: 20000851
DOI: 10.1021/bi901503g

実験手法

X-RAY DIFFRACTION (1.9 Å)