Type A Allatostatins from Drosophila melanogaster and Diplotera puncata Activate Two Drosophila Allatostatin Receptors, DAR-1 and DAR-2, Expressed in CHO Cells
| Autor: | Teresa Kubiak, Marjorie R. Zantello, Martha J. Larsen, Katherine J. Burton, David L Lowery, Valdin G. Smith |
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| Rok vydání: | 2001 |
| Předmět: |
Receptors
Neuropeptide medicine.medical_specialty Time Factors G protein Amino Acid Motifs Biophysics Receptors Cell Surface CHO Cells Biology Ligands Transfection Pertussis toxin Biochemistry Preprohormone Receptors G-Protein-Coupled Cricetinae Internal medicine parasitic diseases medicine Animals Drosophila Proteins Virulence Factors Bordetella Cloning Molecular Receptor Molecular Biology G protein-coupled receptor Dose-Response Relationship Drug Diptera Neuropeptides fungi Allatostatin Cell Biology biology.organism_classification Molecular biology Protein Structure Tertiary Kinetics Drosophila melanogaster Endocrinology Pertussis Toxin Guanosine 5'-O-(3-Thiotriphosphate) Insect Proteins Calcium Signal transduction Protein Binding Signal Transduction |
| Zdroj: | Biochemical and Biophysical Research Communications. 286:895-901 |
| ISSN: | 0006-291X |
| DOI: | 10.1006/bbrc.2001.5476 |
| Popis: | The type-A allatostatins A (AST-A) are a group of insect peptides with a common C-terminal motif Y/FXFGL-NH 2 . The existence of at least four putative type A Drosophila melanogaster ASTs (called type A drostatins or DST-As) has been predicted from the sequence of a recently cloned DST-A preprohormone [C. Lenz et al. (2000) Biochem. Biophys. Res. Commun. 273, 126–1131]. SRPYSFGL-NH 2 , (DST-3A), the only DST isolated from Drosophila so far, activated the first cloned DST-A GPCR (DAR-1) [N. Birgul et al. (1999) EMBO J. 18, 5892–5900]. A newly cloned orphan Dm GPCR, which shares 47% overall and 60% transmembrane region sequence identity with DAR-1, was classified as a second putative Dm DST-A receptor (DAR-2) [C. Lenz et al. (2000) Biochem. Biophys. Res. Commun. 273, 571–577]. Although activation of DAR-2 by DSTs has been postulated, no experimental evidence for that has been presented to date. In this study, we expressed both DAR-1 and DAR-2 in CHO cells and used a GTPγS and a Ca 2+ mobilization assay for pharmacological evaluation of the receptors. Synthetically prepared DST-As, as well as selected Diplotera punctata (cockroach) ASTs, activated DAR-1 and DAR-2 in both functional assays indicating ligand redundancy and cross species activity. Cell pretreatment with pertussis toxin led to some differences in the nature and magnitude of signaling pathways at the DAR-1 and DAR-2 receptors, suggesting possible differential coupling to cellular effector system(s) and distinct biological functions of each receptor in vivo. |
| Databáze: | OpenAIRE |
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