4G4H

100K X-ray diffraction study of carboplatin binding to HEWL in DMSO media after 13 months of crystal storage

4G4H の概要

• エントリーDOI: 10.2210/pdb4g4h/pdb
• 関連するPDBエントリー: 4DD0 4DD1 4DD2 4DD3 4DD4 4DD6 4DD7 4DD9 4DDA 4DDB 4DDC 4G49 4G4A 4G4B 4G4C
• 分子名称: Lysozyme C, DIMETHYL SULFOXIDE, CHLORIDE ION, ... (5 entities in total)
• 機能のキーワード: cisplatin, carboplatin, aqueous media, dmso media, omega scan data collection, capillaries, hydrolase
• 由来する生物種: Gallus gallus (chicken)
• 細胞内の位置: Secreted: P00698
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 15336.28

構造登録者

Tanley, S.W.M.,Schreurs, A.M.M.,Kroon-Batenburg, L.M.J.,Helliwell, J.R. (登録日: 2012-07-16, 公開日: 2012-11-07, 最終更新日: 2023-09-13)

主引用文献

Tanley, S.W.,Schreurs, A.M.,Kroon-Batenburg, L.M.,Helliwell, J.R.
Room-temperature X-ray diffraction studies of cisplatin and carboplatin binding to His15 of HEWL after prolonged chemical exposure.
Acta Crystallogr.,Sect.F, 68:1300-1306, 2012

PubMed Abstract: The anticancer complexes cisplatin and carboplatin are known to bind to both the Nδ and the Nℇ atoms of His15 of hen egg-white lysozyme (HEWL) in the presence of dimethyl sulfoxide (DMSO). However, neither binds in aqueous media after 4 d of crystallization and crystal growth, suggesting that DMSO facilitates cisplatin/carboplatin binding to the N atoms of His15 by an unknown mechanism. Crystals of HEWL cocrystallized with cisplatin in both aqueous and DMSO media, of HEWL cocrystallized with carboplatin in DMSO medium and of HEWL cocrystallized with cisplatin and N-acetylglucosamine (NAG) in DMSO medium were stored for between seven and 15 months. X-ray diffraction studies of these crystals were carried out on a Bruker APEX II home-source diffractometer at room temperature. Room-temperature X-ray diffraction data collection removed the need for cryoprotectants to be used, ruling out any effect that the cryoprotectants might have had on binding to the protein. Both cisplatin and carboplatin still bind to both the Nδ and Nℇ atoms of His15 in DMSO media as expected, but more detail for the cyclobutanedicarboxylate (CBDC) moiety of carboplatin was observed at the Nℇ binding site. However, two molecules of cisplatin were now observed to be bound to His15 in aqueous conditions. The platinum peak positions were identified using anomalous difference electron-density maps as a cross-check with Fo-Fc OMIT electron-density maps. The occupancies of each binding site were calculated using SHELXTL. These results show that over time cisplatin binds to both N atoms of His15 of HEWL in aqueous media, whereas this binding is speeded up in the presence of DMSO. The implication of cisplatin binding to proteins after a prolonged period of time is an important consideration for the length of treatment in patients who are given cisplatin.

PubMed: 23143236
DOI: 10.1107/S1744309112042005

実験手法

X-RAY DIFFRACTION (2 Å)