Isolation and characterization of a novel lectin from the mushroom Armillaria luteo-virens.
From the dried fruiting bodies of the mushroom Armillaria luteo-virens, a dimeric lectin with a molecular mass of 29.4 kDa has been isolated. The purification procedure involved (NH(4))(2)SO(4) precipitation, ion exchange chromatography on DEAE-cellulose, CM-cellulose, and Q-Sepharose, and gel filtration by fast protein liquid chromatography on Superdex 75. The hemagglutinating activity of the lectin could not be inhibited by simple sugars but was inhibited by the polysaccharide inulin. The activity was stable up to 70 degrees C but was acid- and alkali-labile. Salts including FeCl(3), AlCl(3), and ZnCl(2) inhibited the activity whereas MgCl(2), MnCl(2), and CaCl(2) did not. The lectin stimulated mitogenic response of mouse splenocytes with the maximal response achieved by 1microM lectin. Proliferation of tumor cells including MBL2 cells, HeLa cells, and L1210 cells was inhibited by the lectin with an IC(50) of 2.5, 5, and 10 microM, respectively. However, proliferation of HepG2 cells was not affected. The novel aspects of the isolated lectin include a novel N-terminal sequence, fair thermostability, acid stability, and alkali stability, together with potent mitogenic activity toward spleen cells and antiproliferative activity toward tumor cells.
Feng
K
K
State Key Laboratory for Agrobiotechnology and Department of Microbiology, China Agricultural University, Beijing.
Liu
Q H
QH
Ng
T B
TB
Liu
H Z
HZ
Li
J Q
JQ
Chen
G
G
Sheng
H Y
HY
Xie
Z L
ZL
Wang
H X
HX
D000363:Agaricales / Q000737:chemistry*; D000818:Animals; D002241:Carbohydrates / Q000494:pharmacology; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002852:Chromatography, Ion Exchange; D004591:Electrophoresis, Polyacrylamide Gel; D004912:Erythrocytes / Q000187:drug effects / Q000276:immunology; D005656:Fungal Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D006367:HeLa Cells; D006386:Hemagglutination Tests; D006801:Humans; D020128:Inhibitory Concentration 50; D037102:Lectins / Q000737:chemistry / Q000302:isolation & purification* / Q000494:pharmacology; D051379:Mice; D008807:Mice, Inbred BALB C; D008970:Molecular Weight; D011817:Rabbits; D020539:Sequence Analysis, Protein
D002241:Carbohydrates; D005656:Fungal Proteins; D037102:Lectins