Isolation and expression of a sterol carrier protein-2 gene from the yellow fever mosquito, Aedes aegypti.
Trafficking of cholesterol in insects is a very important process due to the fact that insects depend on dietary cholesterol to fulfil their physiological needs. We identified a putative mosquito sterol carrier protein-2 (SCP-2) cDNA from fourth instar subtracted cDNA library. The AeSCP-2 protein has high degree homology in the sterol transfer domain to both rat and human SCP-2. Transcripts of AeSCP-2 in fourth instars were detected strongly in the midgut, and weakly in the head and hindgut. In the early pupae, AeSCP-2 transcription was observed in the thorax, head and body wall of abdomen, but not in the gut. The interaction of mosquito sterol carrier protein-2 (AeSCP-2) with cholesterol was examined. The Kd of purified recombinant AeSCP-2 to cholesterol was 5.6 +/- 0.6 x 10-9 m using radiolabelled cholesterol-binding assay. The results suggest that AeSCP-2 has high affinity to cholesterol and may function as a carrier protein in mosquitoes.
Krebs
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Department of Entomology, University of Wisconsin-Madison, Madison, WI 53076, USA.
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D000330:Aedes / Q000235:genetics* / Q000378:metabolism; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D015152:Blotting, Northern; D015153:Blotting, Western; D002352:Carrier Proteins / Q000235:genetics* / Q000378:metabolism; D002784:Cholesterol / Q000378:metabolism*; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D005260:Female; D018507:Gene Expression Regulation, Developmental; D008969:Molecular Sequence Data; D012313:RNA / Q000737:chemistry / Q000235:genetics; D019105:Random Amplified Polymorphic DNA Technique; D016415:Sequence Alignment
D002352:Carrier Proteins; D018076:DNA, Complementary; D012313:RNA; D002784:Cholesterol