Volf P., kaĝupová S. and Man P., 2002. Characterization of the lectin from females
of Phlebotomus duboscqi sand flies. European Journal of Biochemistry 269(24):
6294-6301.
ISSN 014-2956, IF 2,849
Abstract:
Lectin from females of the important sand fly vector, Phlebotomus duboscqi (Diptera:
Psychodidae), was isolated by immunoaffinity chromatography using a minicolumn
with immobilized anti-lectin immunoglobulins. Carbohydrate-binding specificity
of active fractions corresponded to that of midgut and salivary gland lysates.
Haemagglutination was inhibited by d-glucosamine, d-galactosamine and d-mannosamine.
The homogeneity and molecular mass of the purified lectin was examined by SDS/PAGE
in both reducing and nonreducing conditions. The active fractions showed one band
strongly stained by Coomassie blue or silver nitrate; the molecular mass of the
lectin was 42 kDa under nonreducing and 44 kDa under reducing conditions. SDS/PAGE
of active fractions from the gel filtration revealed four to six protein bands,
but the 42/44-kDa protein present in all active fractions was the only component
reacting with specific antibodies in Western blots. Localization of the lectin
in the gut of females was studied using indirect immunofluorescence on sections.
The positive reaction of specific antibodies was localized in the lumen and along
the microvillar surfaces of epithelial cells. The lectin was partially sequenced
and characterized by MS. Peptide maps were obtained by MALDI-TOF MS, and several
sequence tags were identified from tandem mass spectra on an ion trap. These sequences
displayed high similarity to salivary protein precursors previously identified
in a cDNA library of the sand flies Phlebotomus papatasi and Lutzomyia longipalpis
. Two main hypotheses on the role of female lectin in Leishmania development are
discussed.
Author Keywords:
immunoaffinity chromatography; lectin; Phlebotomus duboscqi; sand fly
CARBOHYDRATE-BINDING SPECIFICITIES; PHYSICOCHEMICAL PROPERTIES; HEMAGGLUTINATION
ACTIVITY; LEISHMANIA; MIDGUT; SANDFLIES; GLOSSINA; INFECTIONS; PAPATASI; VECTOR