Glucose uptake in the mammalian stages of Trypanosoma cruzi.
Trypanosoma cruzi, the agent of Chagas' disease, alternates between different morphogenetic stages that face distinct physiological conditions in their invertebrate and vertebrate hosts, likely in the availability of glucose. While the glucose transport is well characterized in epimastigotes of T. cruzi, nothing is known about how the mammalian stages acquire this molecule. Herein glucose transport activity and expression were analyzed in the three developmental stages present in the vertebrate cycle of T. cruzi. The infective trypomastigotes showed the highest transport activity (V(max)=5.34+/-0.54 nmol/min per mg of protein; K(m)=0.38+/-0.01 mM) when compared to intracellular epimastigotes (V(max)=2.18+/-0.20 nmol/min per mg of protein; K(m)=0.39+/-0.01 mM). Under the conditions employed no transport activity could be detected in amastigotes. The gene of the glucose transporter is expressed at the mRNA level in trypomastigotes and in intracellular epimastigotes but not in amastigotes, as revealed by real-time PCR. In both trypomastigotes and intracellular epimastigotes protein expression could be detected by Western blot with an antibody raised against the glucose transporter correlating well with the transport activity measured experimentally. Interestingly, anti-glucose transporter antibodies showed a strong reactivity with glycosome and reservosome organelles. A comparison between proline and glucose transport among the intracellular differentiation forms is presented. The data suggest that the regulation of glucose transporter reflects different energy and carbon requirements along the intracellular life cycle of T. cruzi.
Silber
Ariel M
AM
Departamento de Bioquímica, Instituto de Química, Universidade de São Paulo, Av. Prof Lineu Prestes 748, São Paulo, SP, Brazil.
Tonelli
Renata R
RR
Lopes
Camila G
CG
Cunha-e-Silva
Narcisa
N
Torrecilhas
Ana Cláudia T
AC
Schumacher
Robert I
RI
Colli
Walter
W
Alves
Maria Júlia M
MJ
D000818:Animals; D015153:Blotting, Western; D020869:Gene Expression Profiling; D005947:Glucose / Q000378:metabolism*; D011392:Proline / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D022161:Transport Vesicles; D014349:Trypanosoma cruzi / Q000378:metabolism*
D011392:Proline; D005947:Glucose