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. 2019 Mar 19:2019:1341370.
doi: 10.1155/2019/1341370. eCollection 2019.

An Inulin-Specific Lectin with Anti-HIV-1 Reverse Transcriptase, Antiproliferative, and Mitogenic Activities from the Edible Mushroom Agaricus bitorquis

Guo-Qing Zhang  1 , Qing-Jun Chen  2 , Jing Hua  1 , Zi-Lu Liu  1 , Yue Sun  1 , Xin Xu  1 , Peng Han  1 , He-Xiang Wang  3
Affiliations

An Inulin-Specific Lectin with Anti-HIV-1 Reverse Transcriptase, Antiproliferative, and Mitogenic Activities from the Edible Mushroom Agaricus bitorquis

Guo-Qing Zhang et al. Biomed Res Int. .

Abstract

A novel lectin (ABL) was purified from the dried fruiting bodies of Agaricus bitorquis. An efficient 3-step purification protocol involved two consecutive steps of ion exchange chromatography on Q-Sepharose and SP-Sepharose and gel filtration by FPLC on Superdex 75. ABL is a monomeric protein with the molecular mass of 27.6 kDa, which is different from other lectins from genus Agaricus. Its N-terminal amino acid sequence is EYTISIRVYQTNPKGFNRPV which is unique and sharing considerably high similarity of other mushroom lectins. The hemagglutinating activity of the lectin was inhibited by inulin. Based on hemagglutination tests, ABL prefers rabbit, human type A, and AB erythrocytes to human type B and O erythrocytes. The lectin inhibits the activity of HIV-1 reverse transcriptase and the proliferation of leukemia cell (L1210) with an IC50 value of 4.69 and 4.97 μM, respectively. Furthermore, ABL demonstrates the highest mitogenic activity with a response of 24177.7 ± 940.6 [3H-methyl] thymidine counts per minute (CPM) at a concentration of 0.91 μM.

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Figures

Figure 1
Figure 1
(a) FPLC-gel filtration on Superdex 75 HR 10/30 column. Eluent: 0.2 M NH4HCO3 buffer (pH 9.4). Fraction size: 0.8 ml. Flow rate: 0.5 ml/min. Fraction SU2 represents purified lectin ABL. (b) SDS-PAGE of ABL (right lane) and molecular weight markers (left lane). The markers were phosphorylase b (94kDa), bovine serum albumin (67kDa), ovalbumin (43kDa), carbonic anhydrase (30 kDa), soybean trypsin inhibitor (20kDa), and α-lactalbumin (14.4kDa), all from GE Healthcare.
Figure 2
Figure 2
HIV-1 reverse transcriptase inhibitory activity of ABL (data represent means ± SD, n = 3).
Figure 3
Figure 3
Inhibitory effect of ABL on proliferation of HepG2 and L1210 cancer cells. Cell proliferation was determined by MTT assay (data represent means ± SD, n = 3).
Figure 4
Figure 4
Mitogenic response of murine splenocytes induced by ABL and ConA (data represent means ± SD, n = 3).
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