Identification of the Neurofibromatosis Type 1 Gene Product

• Published in: Proceedings of the National Academy of Sciences - PNAS 1991-11, Vol.88 (21), p.9658-9662
• Main Authors: Gutmann, David H., Wood, Deborah L., Collins, Francis S.
• Format: Article
• Language: English
• Subjects: 550201 - Biochemistry- Tracer Techniques; Amino Acid Sequence; AMINO ACIDS; Animals; Antibodies; Antiserum; BASIC BIOLOGICAL SCIENCES; BETA DECAY RADIOISOTOPES; BETA-MINUS DECAY RADIOISOTOPES; BIOASSAY; Biological and medical sciences; Blotting, Western; CARBOXYLIC ACIDS; Cell lines; CHROMOSOMAL ABERRATIONS; CHROMOSOMES; Complementary DNA; DAYS LIVING RADIOISOTOPES; DISEASES; DOMINANT MUTATIONS; DRUGS; ELECTROPHORESIS; Enzyme Activation; ENZYME IMMUNOASSAY; Epitopes; ETIOLOGY; EVEN-ODD NUCLEI; Gene Expression; GENES; Genes, Neurofibromatosis 1; GLIOMAS; GTP Phosphohydrolases - metabolism; GTPase-Activating Proteins; guanine nucleotide-binding protein; HEREDITARY DISEASES; HUMAN CHROMOSOME 17; HUMAN CHROMOSOMES; Humans; IMMUNE SERUMS; IMMUNOASSAY; Immunoblotting; Immunoprecipitation; ISOTOPES; LIGHT NUCLEI; LIPOTROPIC FACTORS; Medical sciences; METHIONINE; Mice; Molecular Sequence Data; Molecular Weight; MUTATIONS; NEOPLASMS; Neurofibromatoses; Neurofibromatosis 1; Neurofibromatosis 1 - genetics; Neurofibromin 1; Neurology; NUCLEI; ORGANIC ACIDS; ORGANIC COMPOUNDS; ORGANIC SULFUR COMPOUNDS; PATIENTS; Peptides - chemistry; Peptides - immunology; PROTEINS; Proteins - chemistry; Proteins - genetics; Proteins - immunology; RADIOISOTOPES; Recombinant Fusion Proteins - immunology; SULFUR 35; SULFUR ISOTOPES; TRANSLOCATION; Tumors of the nervous system. Phacomatoses
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.88.21.9658

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 ≈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.