Identification and characterization of porcine NP-190, a novel protein that is specifically expressed in the axonal membrane during the embryonic period.
To identify and analyze the function of proteins expressed in the growth cones, we have screened monoclonal antibodies raised against the preparation of the growth cone particles derived from fetal porcine brains and found a novel neuronal antigen, termed NP-190. Biochemical characterization of NP-190 demonstrated that it was an integral membrane protein with an apparent molecular weight of 190 kDa and that it was mainly expressed in fetal brains. Homologous antigens with molecular weights of 200 and 170 kDa were also identified in the fetal brain extracts of chickens and rats, respectively. Immunoblot experiments of brain extracts from chickens and rats in various stages of development indicated that the expression of NP-190 homologs was developmentally regulated; it began to appear and increased in the embryonic stage, then decreased to very low level in the adult brains. Immunostaining of cultured primary of neurons from the embryonic day 18 rat cerebral cortex demonstrated that rat NP-190 homolog localized in the cell bodies, axons and growth cones, but not in dendrites. Partial amino acid sequence analysis of affinity-purified NP-190 from fetal porcine brains demonstrated that it was a novel protein. These results suggest that NP-190 plays a distinct role in brain development.
Ho
S C
SC
Department of Biotechnology, The University of Tokyo.
Wakatsuki
S
S
Arioka
M
M
Yamasaki
M
M
Kitamoto
K
K
D000595:Amino Acid Sequence; D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D000941:Antigens; D001369:Axons / Q000378:metabolism*; D001921:Brain / Q000196:embryology / Q000378:metabolism; D002462:Cell Membrane / Q000378:metabolism; D002478:Cells, Cultured; D002645:Chickens; D005314:Embryonic and Fetal Development; D008565:Membrane Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000302:isolation & purification; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D009419:Nerve Tissue Proteins / Q000096:biosynthesis / Q000737:chemistry* / Q000276:immunology / Q000302:isolation & purification; D009474:Neurons / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D017421:Sequence Analysis; D017386:Sequence Homology, Amino Acid; D013347:Subcellular Fractions / Q000378:metabolism; D013552:Swine
D000911:Antibodies, Monoclonal; D000941:Antigens; D008565:Membrane Proteins; C113677:NP-190 protein, Sus scrofa; D009419:Nerve Tissue Proteins