A novel protein that binds juvenile hormone esterase in fat body tissue and pericardial cells of the tobacco hornworm Manduca sexta L.
Juvenile hormone esterase degrades juvenile hormone, which acts in conjunction with ecdysteroids to control gene expression in insects. Circulating juvenile hormone esterase is removed from insect blood by pericardial cells and degraded in lysosomes. In experiments designed to characterize proteins involved in the degradation of juvenile hormone esterase, a pericardial cell cDNA phage display library derived from the tobacco hornworm moth Manduca sexta L. was constructed and screened for proteins that bind juvenile hormone esterase. A 732-base pair cDNA encoding a novel 29-kDa protein (P29) was isolated. Western and Northern analyses indicated that P29 is present in both pericardial cell and fat body tissues and is expressed in each larval instar. In immunoprecipitation experiments, P29 bound injected recombinant juvenile hormone esterase taken up by pericardial cells and native M. sexta juvenile hormone esterase in fat body tissue, where the enzyme is synthesized. Binding assays showed that P29 bound juvenile hormone esterase more strongly than it did a mutant form of the enzyme with mutations that perturb lysosomal targeting. Based on these data, we propose that P29 functions in pericardial cells to facilitate lysosomal degradation of juvenile hormone esterase.
Shanmugavelu
M
M
Department of Entomology and the Program in Genetics, Iowa State University, Ames, Iowa 50011, USA.
Baytan
A R
AR
Chesnut
J D
JD
Bonning
B C
BC
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D001710:Biotin / Q000378:metabolism; D002265:Carboxylic Ester Hydrolases / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D005216:Fat Body / Q000201:enzymology*; D015723:Gene Library; D019476:Insect Proteins / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D018402:Manduca / Q000201:enzymology*; D008957:Models, Genetic; D008969:Molecular Sequence Data; D019151:Peptide Library; D010496:Pericardium / Q000201:enzymology*; D011233:Precipitin Tests; D011485:Protein Binding; D011994:Recombinant Proteins / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism
D019476:Insect Proteins; D019151:Peptide Library; D011994:Recombinant Proteins; C404103:juvenile hormone esterase-binding protein, 29-kDa, Manduca sexta; D001710:Biotin; D002265:Carboxylic Ester Hydrolases; C038003:juvenile hormone esterase