Purification and characterization of a novel lectin from the toxic wild mushroom Inocybe umbrinella.
From the dried fruiting bodies of the toxic mushroom Inocybe umbrinella, a novel lectin with a molecular mass of 17 kDa has been isolated with about 160-fold purification. The purification protocol comprised ion exchange chromatography on DEAE-cellulose, and CM-cellulose, and gel filtration on Superdex 75. Among the carbohydrates tested, raffinose, d-melibiose, alpha-lactose and d(+)-galactose could inhibit the hemagglutinating activity of the lectin. The hemagglutinating activity was stable between 10 and 60 degrees C, in 12.5-100mM HCl, and in 50mM NaOH. The hemagglutinating activity was inhibited by Ca(2+), Mn(2+)and Mg(2+) ions, but was unaffected by Fe(3+), Zn(2+) and Al(3+) ions. The lectin inhibited HIV-1 reverse transcriptase with an IC(50) of 4.7+/-0.2 microM. Proliferation of tumor cells including hepatoma HepG2 cells and breast cancer MCF7 cells was inhibited by the lectin with an IC(50) of 3.5+/-0.2 microM and 7.4+/-0.3 micoM, respectively. The lectin has a unique N-terminal amino acid sequence, DGVLATNAVA. It did not exhibit antifungal activity. The present report is the first on an Inocybe lectin and represents one of the very few reports on lectins from toxic mushrooms.
Zhao
J K
JK
State Key Laboratory for Agrobiotechnology and Department of Microbiology, China Agricultural University, Beijing 100094, China.
Wang
H X
HX
Ng
T B
TB
D000363:Agaricales / Q000737:chemistry*; D000595:Amino Acid Sequence; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D054303:HIV Reverse Transcriptase / Q000037:antagonists & inhibitors; D006386:Hemagglutination Tests; D006801:Humans; D020128:Inhibitory Concentration 50; D037102:Lectins / Q000235:genetics / Q000302:isolation & purification* / Q000494:pharmacology
D037102:Lectins; D054303:HIV Reverse Transcriptase