Purification and characterization of a new D-galactose-specific lectin from the housefly, Musca domestica, and its antiproliferative effect on human K562 and MCF-7 tumor cells.
In the present work, a D-galactose-specific lectin with novel N-terminal sequence was purified from Musca domestica L. (Diptera: Muscidae) pupae. The purification was performed using affinity chromatography, ultra-filtration, and HPLC. The haemagglutinating activity of M. domestica lectin was specifically inhibited by D-galactose. The haemagglutinating activity of this lectin was stable at temperatures up to 65 degrees C and in pH ranging from 4 to 8. Salts including FeCl(3) and MnCl(2) inhibited the haemagglutinating process, whereas NaCl, KCl, CaCl(2), MgCl(2), ZnCl(2), and AlCl(3) did not. By SDS-PAGE, purified M. domestica pupae lectin yielded a single band with a molecular weight of 40 kDa, with or without reduction of beta-mercaptoethanol, and it could be stained with Alcian Blue 8 GX. The morphology of purified lectin was observed by atomic force microscopy, which indicated that M. domestica lectin was an 8.27 nm high, globular shaped glycoprotein with a 1.41 nm high polysaccharide chain. In addition, antiproliferative activity of this lectin against tumor cells K562 and MCF-7 was determined with a colorimetric assay using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide, which showed that the antiproliferative process was time- and dose-dependent with an IC(50) of 5.7 and 6.7 at 24 h, 5.5 and 6.4 at 36 h, 5.2 and 6.5 microM at 48 h, respectively.
Cao
X
X
Tianjin University of Science and Technology, Tianjin, P.R. China. caoxh@tust.edu.cn
Sun
Y
Y
Wang
C
C
Zeng
B
B
D000818:Animals; D000970:Antineoplastic Agents / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology*; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D005690:Galactose / Q000378:metabolism; D006384:Hemagglutination / Q000187:drug effects; D006793:Houseflies / Q000737:chemistry*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007477:Ions / Q000494:pharmacology; D020014:K562 Cells; D037102:Lectins / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology*; D008670:Metals / Q000494:pharmacology; D013696:Temperature
D000970:Antineoplastic Agents; D007477:Ions; D037102:Lectins; D008670:Metals; D005690:Galactose