Characterization of lipophorin binding to the fat body of Rhodnius prolixus.
In insects, lipids are transported by a hemolymphatic lipoprotein, lipophorin. The binding of lipophorin to the fat body of the hematophagous insect Rhodnius prolixus was characterized in a fat body membrane preparation, obtained from adult females. For the binding assay, purified lipophorin was radiolabelled in the protein moiety (125I-HDLp), and it was shown that iodination did not affect the affinity of the membrane preparation for lipophorin. Under incubation conditions used, lipophorin binding to membranes achieved equilibrium after 40-60 min, but this time was longer when a low concentration of lipophorin was present in the medium. The capacity of the fat body membrane preparation to bind lipophorin was abolished when membranes were pre-treated with trypsin, and it was also affected by heat. When 125I-HDLp was incubated with increasing concentrations of membrane protein, corresponding increases in binding were observed. Lipophorin binding was sensitive to pH, and it was maximal between pH 6.0 and 7.0. The specific binding of lipophorin to the fat body membrane preparation was a saturable process, with a Kd of 2.1 +/- 0.4 x 10(-7)M and a maximal binding capacity of 289 +/- 88 ng lipophorin/microgram of membrane protein. Binding to the fat body membranes did not depend on calcium, but it was affected by ionic strength, being totally inhibited at high salt concentrations. Suramin also interfered with lipophorin binding and it was abolished in the presence of 2 mM suramin, but at concentrations of 0.05 and 0.1 mM it seemed to increase binding activity slightly. Fat body membrane preparation from Rhodnius prolixus was able to bind lipophorin from Manduca sexta larvae.
Pontes
Emerson G
EG
Departamento de Bioquímica Médica, Instituto de Ciências Biomédicas, Universidade Federal do Rio de Janeiro, Rio de Janeiro, 21941-590, Brazil.
Grillo
Luciano A M
LA
Gondim
Katia C
KC
D000273:Adipose Tissue / Q000378:metabolism*; D000818:Animals; D002352:Carrier Proteins / Q000302:isolation & purification / Q000378:metabolism*; D002462:Cell Membrane / Q000378:metabolism; D005260:Female; D006863:Hydrogen-Ion Concentration; D007457:Iodine Radioisotopes; D007700:Kinetics; D050356:Lipid Metabolism; D008074:Lipoproteins / Q000302:isolation & purification / Q000378:metabolism*; D012239:Rhodnius / Q000378:metabolism*; D013816:Thermodynamics; D014357:Trypsin / Q000378:metabolism
D002352:Carrier Proteins; D007457:Iodine Radioisotopes; D008074:Lipoproteins; C032830:lipophorin; D014357:Trypsin