4MX9

CDPK1 from Neospora caninum in complex with inhibitor UW1294

4MX9 の概要

• エントリーDOI: 10.2210/pdb4mx9/pdb
• 関連するPDBエントリー: 4M97
• 分子名称: Calmodulin-like domain protein kinase isoenzyme gamma, related, 3-(6-ethoxynaphthalen-2-yl)-1-[(1-methylpiperidin-4-yl)methyl]-1H-pyrazolo[3,4-d]pyrimidin-4-amine (3 entities in total)
• 機能のキーワード: serine/threonine protein kinase, transferase, calcium-binding, atp-binding, transferase-transferase inhibitor complex, transferase/transferase inhibitor
• 由来する生物種: Neospora caninum
• タンパク質・核酸の鎖数: 1
• 化学式量合計: 55500.21

構造登録者

Merritt, E.A. (登録日: 2013-09-26, 公開日: 2013-10-09, 最終更新日: 2023-09-20)

主引用文献

Ojo, K.K.,Reid, M.C.,Kallur Siddaramaiah, L.,Muller, J.,Winzer, P.,Zhang, Z.,Keyloun, K.R.,Vidadala, R.S.,Merritt, E.A.,Hol, W.G.,Maly, D.J.,Fan, E.,Van Voorhis, W.C.,Hemphill, A.
Neospora caninum Calcium-Dependent Protein Kinase 1 Is an Effective Drug Target for Neosporosis Therapy.
Plos One, 9:e92929-e92929, 2014

PubMed Abstract: Despite the enormous economic importance of Neospora caninum related veterinary diseases, the number of effective therapeutic agents is relatively small. Development of new therapeutic strategies to combat the economic impact of neosporosis remains an important scientific endeavor. This study demonstrates molecular, structural and phenotypic evidence that N. caninum calcium-dependent protein kinase 1 (NcCDPK1) is a promising molecular target for neosporosis drug development. Recombinant NcCDPK1 was expressed, purified and screened against a select group of bumped kinase inhibitors (BKIs) previously shown to have low IC50s against Toxoplasma gondii CDPK1 and T. gondii tachyzoites. NcCDPK1 was inhibited by low concentrations of BKIs. The three-dimensional structure of NcCDPK1 in complex with BKIs was studied crystallographically. The BKI-NcCDPK1 structures demonstrated the structural basis for potency and selectivity. Calcium-dependent conformational changes in solution as characterized by small-angle X-ray scattering are consistent with previous structures in low Calcium-state but different in the Calcium-bound active state than predicted by X-ray crystallography. BKIs effectively inhibited N. caninum tachyzoite proliferation in vitro. Electron microscopic analysis of N. caninum cells revealed ultra-structural changes in the presence of BKI compound 1294. BKI compound 1294 interfered with an early step in Neospora tachyzoite host cell invasion and egress. Prolonged incubation in the presence of 1294 interfered produced observable interference with viability and replication. Oral dosing of BKI compound 1294 at 50 mg/kg for 5 days in established murine neosporosis resulted in a 10-fold reduced cerebral parasite burden compared to untreated control. Further experiments are needed to determine the PK, optimal dosage, and duration for effective treatment in cattle and dogs, but these data demonstrate proof-of-concept for BKIs, and 1294 specifically, for therapy of bovine and canine neosporosis.

PubMed: 24681759
DOI: 10.1371/journal.pone.0092929

実験手法

X-RAY DIFFRACTION (3.1 Å)