Biochemical characterization of serine transport in Leishmania (Leishmania) amazonensis.
In addition to its role as a protein component in Leishmania, serine is also a precursor for the synthesis of both phosphatidylserine, which is a membrane molecule involved in parasite invasion and inactivation of macrophages, and sphingolipids, which are necessary for Leishmania to differentiate into its infective forms. We have characterized serine uptake in both promastigote and amastigote forms of Leishmania (Leishmania) amazonensis. In promastigotes, kinetic data show a single, saturable transport system, with a Km of 0.253+/-0.01 mM and a maximum velocity of 0.246+/-0.04 nmol/min per 10(7) cells. Serine transport increased linearly with temperature in the range from 20 degrees C to 45 degrees C, allowing the calculation of an activation energy of 7.09 kJ/mol. Alanine, cysteine, glycine, threonine, valine and ethanolamine competed with the substrate at a ten-fold excess concentration. Serine uptake was dependent on pH, with an optimum activity at pH 7.5. The characterization of the serine transport process in amastigotes revealed a transport system with a similar Km, energy of activation and pH response to that found in promastigotes, suggesting that the same transport system is active in both insect vector and mammalian host Leishmania stages. This could constitute an evolutionary mechanism that guarantees the provision of such an essential molecule during host change events, such as differentiation into amastigotes and macrophage invasion, as well as to ensure that the parasite maintains the infection in the mammalian host.
dos Santos
Marcos Gonzaga
MG
Departamento de Fisiologia, Instituto de Biociências, Universidade de São Paulo, São Paulo, SP, Brazil.
Paes
Lisvane Silva
LS
Zampieri
Ricardo Andrade
RA
da Silva
Maria Fernanda Laranjeira
MF
Silber
Ariel Mariano
AM
Floeter-Winter
Lucile Maria
LM
D000818:Animals; D001692:Biological Transport; D006863:Hydrogen-Ion Concentration; D007700:Kinetics; D007891:Leishmania / Q000378:metabolism*; D012694:Serine / Q000378:metabolism*
D012694:Serine