Isolation and identification of a cAMP generating peptide from the flesh fly, Neobellieria bullata (Diptera: Sarcophagidae).
The Manduca sexta Malpighian tubule assay system, developed to monitor adenylate cyclase activity, was used in combination with HPLC to isolate a novel cAMP generating peptide from 350,000 whole flesh flies, Neobellieria bullata. Mass spectrometry revealed a molecular mass of 5,047 daltons, and Edman degradation the following sequence: AGAEAEKLSGLSKYFNGTTMAGRANVAKATYAVIGLIIAYNVMKPKKK. This 48-mer peptide, called Neb-cGP, does not belong to the corticotropin releasing factor family of insect diuretic peptides. Electrophoresis and subsequent immunoblotting of peptides immunoprecipitated from a homogenate of entire flies showed that one fly contained approximately 0.003 to 0.03 micrograms Neb-cGP and that 10 micrograms represents the lowest immunostainable amount on a Western blot.
Spittaels
K
K
Zoologisch Institut, Katholieke Universiteit Leuven, Belgium.
Devreese
B
B
Schoofs
L
L
Neven
H
H
Janssen
I
I
Grauwels
L
L
Van Beeumen
J
J
De Loof
A
A
D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies; D002851:Chromatography, High Pressure Liquid; D000242:Cyclic AMP / Q000096:biosynthesis*; D004175:Diptera / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D015151:Immunoblotting; D019476:Insect Proteins; D008297:Male; D008317:Malpighian Tubules / Q000378:metabolism; D018402:Manduca / Q000378:metabolism*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D012680:Sensitivity and Specificity
D000906:Antibodies; D019476:Insect Proteins; D011506:Proteins; C098180:cAMP generating peptide, flesh fly; D000242:Cyclic AMP