Toxoplasma gondii calcium-dependent protein kinase 1 is a target for selective kinase inhibitors.
New drugs are needed to treat toxoplasmosis. Toxoplasma gondii calcium-dependent protein kinases (TgCDPKs) are attractive targets because they are absent in mammals. We show that TgCDPK1 is inhibited by low nanomolar levels of bumped kinase inhibitors (BKIs), compounds inactive against mammalian kinases. Cocrystal structures of TgCDPK1 with BKIs confirm that the structural basis for selectivity is due to the unique glycine gatekeeper residue in the ATP-binding site. We show that BKIs interfere with an early step in T. gondii infection of human cells in culture. Furthermore, we show that TgCDPK1 is the in vivo target of BKIs because T. gondii expressing a glycine to methionine gatekeeper mutant enzyme show significantly decreased sensitivity to BKIs. Thus, design of selective TgCDPK1 inhibitors with low host toxicity may be achievable.
Ojo
Kayode K
KK
Division of Allergy and Infectious Diseases, Department of Medicine, University of Washington, Seattle, Washington, USA.
Larson
Eric T
ET
Keyloun
Katelyn R
KR
Castaneda
Lisa J
LJ
Derocher
Amy E
AE
Inampudi
Krishna K
KK
Kim
Jessica E
JE
Arakaki
Tracy L
TL
Murphy
Ryan C
RC
Zhang
Li
L
Napuli
Alberto J
AJ
Maly
Dustin J
DJ
Verlinde
Christophe L M J
CL
Buckner
Frederick S
FS
Parsons
Marilyn
M
Hol
Wim G J
WG
Merritt
Ethan A
EA
Van Voorhis
Wesley C
WC
D000595:Amino Acid Sequence; D000818:Animals; D000977:Antiparasitic Agents / Q000737:chemistry / Q000494:pharmacology*; D018360:Crystallography, X-Ray; D005347:Fibroblasts / Q000469:parasitology; D006790:Host-Parasite Interactions / Q000187:drug effects; D006801:Humans; D008969:Molecular Sequence Data; D011487:Protein Conformation; D047428:Protein Kinase Inhibitors / Q000737:chemistry / Q000494:pharmacology*; D011494:Protein Kinases / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism*; D014122:Toxoplasma / Q000166:cytology / Q000187:drug effects / Q000201:enzymology* / Q000254:growth & development; D014123:Toxoplasmosis / Q000188:drug therapy*; D017871:Calcium-Calmodulin-Dependent Protein Kinases
D000977:Antiparasitic Agents; D047428:Protein Kinase Inhibitors; D011494:Protein Kinases; D017871:Calcium-Calmodulin-Dependent Protein Kinases