用噬菌体展示技术分析G_CSF受体的配体结合位点_武婕.pdf

# 246 # Guangdong Medical ournal Mar . 2003, Vol. 24, No . 3 G- CSF * 1 1 1 1 2 2 武 婕 张宏斌 王 捷 郑文岭 王清明 陈惠鹏 1 广州军区广州总医院医学实验科( 广州510010) ; 2 北京放射医学研究所( 100850) = 分析粒细胞集落刺激因子( G- CSF) 受体中参与 体结合的关键性氨基酸残基 以G- CSF 为靶分子, 对随机噬菌体文库进行筛选, 阳性噬菌体克隆进行序列测定并与G- CSF 受体进行同源性比较和分析 得到 一个保守性的序列模式SXXRVX, 其中第4 位氨基酸是高度保守的Arg, 表明Arg 在与G- CSF 的结合中可能起 重要作用将此模式与G - CSF 受体进行同源性比较和序列分析, 发现G- CSF 受体中第288 位的Arg 也是参与 体结 合的关键性氨基酸 模拟小肽和G- CSF 受体中高度保守的Arg 的存在, 说明Arg 在G- CSF 受体与其 体的结 合中起重要作用 = 粒细胞集落刺激因子受体 噬菌体展示技术 作用机制 Identification of residue critical to the binding activity of G- CSF- R using phage display Wu Jie, Zhang H ong bin , Wang Jie, et al . General Hosp ital of Guangzhou Command of PLA , Guangzhou 510010 = Abstract Objective Phage d splay w as used to dent fy res due cr t cal for the b nd ng act v ty of human granulocyte colony - factor receptor( G- CSF- R) . Methods Aff n ty select ve method was used to screen the l brary. The pos t ve phage clone w as se- quenced and compared w th the sequence of G - CSF - R. Results The conserved pept de mot f -SXXRVX. was found. Of these res dues, Arg4 was the h ghly conserved and appeared to be the most mportant for G- CSF b nd ng, n agreement w th the Arg288 of the G - CSF receptor also be ng the cr t cal res due to the b nd ng G- CSF. The pept de could m m c the structure of G - CSF- R to nter- act w th G- CSF. The h ghly conserved Arg probably made an electrostat c nteract on w th Glu19 . The results proved that Arg288 n Fc - Gc loop has the greatest eff ect for the b nd ng act v ty aga n and was helpful to nd cate the mechan sm of nteract on betw een G- CSF and ts receptor. Conclusion The conserved Arg nd cate that t maybe play a key role n G- CSF receptor b