Culekinin depolarizing peptide: a mosquito leucokinin-like peptide that influences insect Malpighian tubule ion transport.
A peptide termed culekinin depolarizing peptide (CDP) was isolated from approximately 1.2 million mosquitos (94% Culex salinarius). The peptide was isolated on the basis of a rapid myotropic assay that utilized a hindgut preparation from Leucophaea maderae and a transepithelial voltage assay that used mosquito Malpighian tubules from Aedes aegypti. A 15% trifluoroacetic acid extraction from the mosquitos, two solid phase extraction steps, and six HPLC steps resulted in the isolation of 9.7 nmol of CDP. This value corresponds to approximately 8 fmol/mosquito. Edman degradation indicated the following sequence for CDP: Asn-Pro-Phe-His-Ser-Trp-Gly-NH2. The sequence was confirmed as the suspected C-terminal amide form of the peptide, since native and synthetic CDP had identical chemical and biological properties. CDP is a member of the leucokinin family of neuropeptides. The leucokinins have been found in three other insect species (Leucophaea maderae, Acheta domesticus and Locusta migratoria) where these peptides were isolated by their myotropic properties alone. CDP shares a C-terminal sequence homology (i.e., Phe-X-Ser-Trp-Gly-NH2) with the rest of the leucokinins. CDP corresponds to the strongest tubule depolarizing activity in the C. salinarius extract. These findings agree with previous structure-activity studies that suggest that mosquitos would contain a leucokinin-like factor that had Phe-His-Ser-Trp-Gly-NH2 as the C-terminal pentapeptide. This is the first leucokinin isolated from blood feeding or holometabolous insects.
Hayes
T K
TK
Laboratory for Invertebrate Neuroendocrine Research, Texas A&M University, College Station 77843.
Holman
G M
GM
Pannabecker
T L
TL
Wright
M S
MS
Strey
A A
AA
Nachman
R J
RJ
Hoel
D F
DF
Olson
J K
JK
Beyenbach
K W
KW
D000330:Aedes / Q000502:physiology*; D000595:Amino Acid Sequence; D000818:Animals; D003058:Cockroaches / Q000502:physiology*; D007422:Intestines; D017136:Ion Transport / Q000502:physiology; D008317:Malpighian Tubules / Q000378:metabolism*; D008969:Molecular Sequence Data; D009119:Muscle Contraction / Q000502:physiology; D009479:Neuropeptides / Q000302:isolation & purification / Q000502:physiology*
D009479:Neuropeptides; C086282:culekinin depolarizing peptide II