TY - JOUR
T1 - Dematin, a human erythrocyte cytoskeletal protein, is a substrate for a recombinant FIKK kinase from Plasmodium falciparum
AU - Brandt, Gabriel S.
AU - Bailey, Scott
N1 - Funding Information:
We are grateful to our colleagues who provided materials, namely Dr. Jürgen Bosch, Dr. Sean Prigge, Dr. Gregory Bowman, the JHMRI Parasite Core facility (Johns Hopkins University) and Dr. James McKnight (Boston University). Dr. Maroya Spalding and Dr. Mary Katherine Tarrant offered indispendable technical advice. MS/MS analysis was performed by the Taplin Center for Biological Mass Spectrometry, Harvard Medical School. This work was supported by the Bloomberg Family Foundation and a Johns Hopkins Malaria Research Institute Postdoctoral Fellowship (to G.S.B.) and Pilot Grant (to S.B.). The funding institution played no role in study design; in the collection, analysis and interpretation of data; in the writing of the report; or in the decision to submit the article for publication.
PY - 2013
Y1 - 2013
N2 - P. falciparum causes the most deadly form of malaria, resulting from the adherence of infected red blood cells to blood vessels. During the blood stage of infection, the parasite secretes a large number of proteins into the host erythrocyte. The secretion of a 20-member family of protein kinases known as FIKK kinases, after a conserved Phe-Ile-Lys-Lys sequence motif, is unique to P. falciparum. Identification of physiological substrates of these kinases may provide perspective on the importance of FIKK kinase activity to P. falciparum virulence. We demonstrate, for the first time, the heterologous expression and purification of a FIKK kinase (PfFk4.1, PFD1165w). The recombinant kinase is active against general substrates and phosphorylates itself. Having demonstrated kinase activity, we incubated recombinant Fk4.1 with parasite and human erythrocyte lysates. No parasite-derived substrates were identified. However, treatment of erythrocyte ghosts shows that the FIKK kinase Fk4.1 phosphorylates dematin, a cytoskeletal protein found at the red blood cell spectrin-actin junction.
AB - P. falciparum causes the most deadly form of malaria, resulting from the adherence of infected red blood cells to blood vessels. During the blood stage of infection, the parasite secretes a large number of proteins into the host erythrocyte. The secretion of a 20-member family of protein kinases known as FIKK kinases, after a conserved Phe-Ile-Lys-Lys sequence motif, is unique to P. falciparum. Identification of physiological substrates of these kinases may provide perspective on the importance of FIKK kinase activity to P. falciparum virulence. We demonstrate, for the first time, the heterologous expression and purification of a FIKK kinase (PfFk4.1, PFD1165w). The recombinant kinase is active against general substrates and phosphorylates itself. Having demonstrated kinase activity, we incubated recombinant Fk4.1 with parasite and human erythrocyte lysates. No parasite-derived substrates were identified. However, treatment of erythrocyte ghosts shows that the FIKK kinase Fk4.1 phosphorylates dematin, a cytoskeletal protein found at the red blood cell spectrin-actin junction.
KW - Autophosphorylation
KW - Dematin
KW - FIKK kinase
KW - Malaria
KW - Pathogen kinase
KW - Plasmodium falciparum
UR - http://www.scopus.com/inward/record.url?scp=84883449074&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84883449074&partnerID=8YFLogxK
U2 - 10.1016/j.molbiopara.2013.08.003
DO - 10.1016/j.molbiopara.2013.08.003
M3 - Article
C2 - 23973789
AN - SCOPUS:84883449074
SN - 0166-6851
VL - 191
SP - 20
EP - 23
JO - Molecular and Biochemical Parasitology
JF - Molecular and Biochemical Parasitology
IS - 1
ER -