Comparison of the protein kinase and acid phosphatase activities of five species of Leishmania.
Promastigotes from log phase and stationary phase cultures of Leishmania donovani, L. braziliensis panamensis, L. tropica, L. major, and L. mexicana amazonensis were analyzed for their content of protein kinase and acid phosphatase activities. Cell surface, histone-specific protein kinase activity was 1.3- to 2.8-fold higher in stationary phase cells of all species except for L. tropica in which the activities of stationary and log phase cells were equal; L. mexicana amazonensis had the highest histone-specific protein kinase activity and L. donovani the lowest. When viable, motile promastigotes of all five species were incubated for 10 min with [gamma-32P]ATP and Mg2+ (10 mM) in the absence of exogenous histone acceptor; about one dozen proteins were phosphorylated in each case. Both log phase and stationary phase promastigotes of all five species extensively phosphorylated a 50-kDa protein that had the mobility of tubulin. Incubation of pure calf brain tubulin with [gamma-32P]ATP and purified L. donovani protein kinase resulted in extensive phosphorylation of the former. Highly infective metacyclic forms (PNA-) of L. major, isolated from a stationary culture using the peanut agglutinin (PNA), contained eight times more histone-specific protein kinase activity than noninfective log phase cells (PNA+). The PNA- and PNA+ forms of L. major both phosphorylated a 50-kDa protein when incubated with [gamma-32P]ATP and magnesium or manganese ions (10 mM); the 50-kDa protein was precipitated by anti-tubulin rabbit antibodies. Extracts of all five species contained large amounts of acid phosphatase activity.(ABSTRACT TRUNCATED AT 250 WORDS)
Mukhopadhyay
N K
NK
Department of Microbiology, School of Medicine, University of Pittsburgh, Pennsylvania 15261.
Saha
A K
AK
Lovelace
J K
JK
Da Silva
R
R
Sacks
D L
DL
Glew
R H
RH
D000135:Acid Phosphatase / Q000378:metabolism*; D000818:Animals; D050260:Carbohydrate Metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D037102:Lectins; D007891:Leishmania / Q000201:enzymology* / Q000472:pathogenicity; D008565:Membrane Proteins / Q000378:metabolism; D019887:Peanut Agglutinin; D010750:Phosphoproteins / Q000378:metabolism; D010766:Phosphorylation; D011494:Protein Kinases / Q000378:metabolism*; D013045:Species Specificity; D014404:Tubulin / Q000378:metabolism
D037102:Lectins; D008565:Membrane Proteins; D019887:Peanut Agglutinin; D010750:Phosphoproteins; D014404:Tubulin; D011494:Protein Kinases; D000135:Acid Phosphatase