title: Structural diversity of trypsin from different mosquito species feeding on vertebrate blood.
abstract: Mosquito trypsin was purified using a combination of ion exchange and affinity chromatography with the ligand soybean trypsin inhibitor. Three Aedes and three Anopheles species were tested, all of which are specialized in the digestion of vertebrate blood. Amino-terminal sequences of HPLC-purified trypsins from Aedes aegypti and Anopheles quadrimaculatus revealed homologies of 30-40% with vertebrate and other invertebrate proteases previously identified as serine-proteases. The purified mosquito trypsins have molecular masses between 25 kDa and 36 kDa, as determined by denaturing polyacrylamide electrophoresis, and are heterogeneous in size and number in the various species. The number of SDS-bands varies between 3 and 6 in Aedes and between 1 and 3 in Anopheles. The specific activities, determined with the substrate TAME, range from 240 U/mg in Aedes aegypti to 1065 U/mg in Anopheles quadrimaculatus. All mosquito trypsins tested have acidic isoelectric points between pH 3.5 and pH 5.4. No alkaline proteases were detected. Polyclonal antisera against Aedes aegypti and Anopheles albimanus trypsin do not cross-react with bovine trypsin. Cross-reactivity of the two sera with trypsin from six mosquito species suggests the presence of at least 2 enzyme families.
authors.affiliation: Zoologisches Institut, Universität Zürich, Switzerland.
authors.forename: R
authors.initials: R
authors.lastname: Graf
authors.forename: P
authors.initials: P
authors.lastname: Boehlen
authors.forename: H
authors.initials: H
authors.lastname: Briegel
mesh_terms: D000330:Aedes / Q000201:enzymology*; D000595:Amino Acid Sequence; D000818:Animals; D000852:Anopheles / Q000201:enzymology*; D001769:Blood / Q000378:metabolism; D015153:Blotting, Western; D002845:Chromatography; D002851:Chromatography, High Pressure Liquid; D007526:Isoelectric Point; D008969:Molecular Sequence Data; D008970:Molecular Weight; D012689:Sequence Homology, Nucleic Acid; D013045:Species Specificity; D014357:Trypsin / Q000737:chemistry* / Q000302:isolation & purification
chemical_list: D014357:Trypsin