灵芝糖蛋白与人源蛋白RANKL胞外结构域的相互作用

Studies on the Interaction Between Extracellular Domain of Human RANKL with Bioactive Glycoprotein

采用酶辅助超声提取法,利用DEAE-Sephadex A25离子层析分离得到一种天然灵芝糖蛋白GP2。经SDS-PAGE检测,以高碘酸希夫试剂染色得到单一条带,说明此组分为单一糖蛋白,红外检测分析证实此组分(GP2)为糖蛋白。荧光检测结果表明,此糖蛋白导致人源RANKL胞外结构域(eRANKL)内源荧光减弱,ANS荧光增强,推测此组分可能会使该蛋白构象发生改变。

Osteoprotegerin （OPG） and receptor activator of nuclear factor kappa B （RANK） are members of the TNFRsuperfamily that regulate osteoclast formation and function by competing for RANK ligand （RANKL）. RANKL promotesosteoclast development through RANK activation, while OPG inhibits this process by sequestering RANKL. In this paper, anatural ganoderma glycoprotein was harvested by the means of ultrasonic assisted, enzymatic action and DEAE - Sephadex A25ion chromatography. The homogeneous glycoprotein from ganoderma was identified by SDS - PAGE, stained with Acid - Schiffregent and infrared analysis. Fluorescence analysis indicated that the glycoprotein could result eRANKL intrinsic fluorescenceintensity dropping and ANS fluorescence intensity increasing, which suggested the glycoprotein caused the eRANKL＇ sconformational change.