Mechanistic studies of the degradation of juvenile hormone esterase in Manduca sexta.
The mechanisms of degradation of juvenile hormone esterase (JHE) were investigated in larvae of the tobacco hornworm, Manduca sexta. JHE is removed from the hemolymph by the pericardial cells by receptor-mediated endocytosis and is ultimately degraded in the lysosomes. Immunoprecipitation experiments and native PAGE followed by Western blotting showed that JHE associates with a putative heat shock cognate protein (Hsp). Approximately 25% of the active JHE in the pericardial cell complex is associated with the putative Hsp 1 h postinjection of affinity purified JHE. Electron microscope analysis revealed that the putative Hsp is located in the trans-Golgi network of pericardial cells, where it is hypothesized to be involved in sorting of proteins destined for the lysosomes, from those destined for the cell membrane. Data acquired from immunoprecipitation and Western blotting experiments argue against the involvement of ubiquitin in the degradation of JHE. Injection of radiolabeled JHE into larvae of M. sexta followed by SDS-PAGE of pericardial cell homogenates revealed covalent binding of an unidentified protein to JHE in the pericardial cell complex.
Bonning
B C
BC
Department of Entomology, Iowa State University, Ames, USA.
Booth
T F
TF
Hammock
B D
BD
D000818:Animals; D002265:Carboxylic Ester Hydrolases / Q000302:isolation & purification / Q000378:metabolism* / Q000648:ultrastructure; D004591:Electrophoresis, Polyacrylamide Gel; D006360:Heat-Shock Proteins / Q000302:isolation & purification / Q000378:metabolism / Q000648:ultrastructure; D006458:Hemolymph / Q000201:enzymology; D008247:Lysosomes / Q000201:enzymology; D018402:Manduca / Q000201:enzymology*; D008854:Microscopy, Electron; D008970:Molecular Weight; D011994:Recombinant Proteins / Q000378:metabolism
D006360:Heat-Shock Proteins; D011994:Recombinant Proteins; D002265:Carboxylic Ester Hydrolases; C038003:juvenile hormone esterase