IDENTIFICATION OF THE NEUROFIBROMATOSIS TYPE-1 GENE-PRODUCT
IDENTIFICATION OF THE NEUROFIBROMATOSIS TYPE-1 GENE-PRODUCT
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DOI：
10.1073/pnas.88.21.9658
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发表时间：
1991-11-01
影响因子：
11.1
通讯作者：
COLLINS, FS
中科院分区：
文献类型：
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作者：
GUTMANN, DH;WOOD, DL;COLLINS, FS
1型神经纤维瘤病（NF1）的基因最近被定位克隆鉴定。完整的cDNA编码2818个氨基酸的多肽。为了研究NF1基因产物，对融合蛋白和合成肽都提出了抗体。两种抗肽抗体和一种融合蛋白抗体的初步鉴定表明，免疫沉淀和免疫印迹的特异性蛋白几乎等于250 kDa。这种蛋白存在于所有组织和细胞系中，在人类、大鼠和小鼠的组织中也能检测到。为了证明这些抗体特异性识别NF1蛋白，产生了含有合成肽特异性序列的额外融合蛋白。两种肽抗血清都能识别相应的特异性融合蛋白。使用肽抗血清的免疫沉淀被证明可以识别与其他肽抗血清或融合蛋白抗血清免疫印迹检测到的相同的蛋白质。在组织匀浆上使用对空间上不同表位特异的抗血清进行免疫印迹检测，结果显示NF1蛋白在所有成人组织中均存在。基于NF1基因产物与gtpase - activated protein （GAP）超家族成员的同源性，建议命名为NF1-GAP相关蛋白（NF1GRP）。
The gene for neurofibromatosis type 1 (NF1) was recently identified by positional cloning. The complete cDNA encodes a polypeptide of 2818 amino acids. To study the NF1 gene product, antibodies were raised against both fusion proteins and synthetic peptides. Initial characterization of two anti-peptide antibodies and one fusion-protein antibody demonstrated a specific protein of almost-equal-to 250 kDa by both immunoprecipitation and immunoblotting. This protein was found in all tissues and cell lines examined and is detected in human, rat, and mouse tissues. To demonstrate that these antibodies specifically recognize the NF1 protein, additional fusion proteins containing the sequence specific to the synthetic peptide were generated. Both peptide antisera recognize the proper specific fusion proteins so generated. Immunoprecipitates using the peptide antisera were shown to recognize the same protein detected by immunoblotting with either the other peptide antiserum or the fusion-protein antiserum. Immunoblotting using antiserum specific to spatially distinct epitopes conducted on tissue homogenates demonstrated the NF1 protein in all adult tissues. Based on the homology between the NF1 gene product and members of the GTPase-activating protein (GAP) superfamily, the name NF1-GAP-related protein (NF1GRP) is suggested.