Berger E A, Shooter E M
J Biol Chem. 1978 Feb 10;253(3):804-10.
The biosynthesis of beta nerve growth factor (betaNGF) was studied in isolated mouse submaxillary glands incubated with L-[35S]cystine. Sodium dodecyl sulfate gels of anti-betaNGF immunoprecipitates from labeled gland homogenates showed a single major peak of radioactivity, which comigrated with purified betaNGF. This species was nearly completely precipitated by the addition of equivalent amounts of anti-betaNGF, but was absent from immunoprecipitates obtained by the addition of ferritin plus anti-ferritin. The cystine-containing tryptic peptides of the labeled species appeared identical with those of purified betaNGF. In submaxillary glands from adult male mice, labeling of betaNGF represented approximately 0.2% of the trichloroacetic acid-precipitable radioactivity. Castration reduced this value to one-third, while testosterone treatment of castrated animals restored the relative betaNGF synthesis to normal or more. No betaNGF synthesis could be detected in glands from female animals. Several tissues were examined for their ability to synthesize betaNGF in culture. Only submaxillary gland incorporated detectable amounts of radioactivity into betaNGF. Labeling of betaNGF could also be obtained by direct injection of isotope into the submaxillary gland in vivo. The results are discussed in terms of the integration of betaNGF synthesis into neuronal development and maintenance.
在与L-[35S]胱氨酸一起孵育的离体小鼠下颌下腺中研究了β神经生长因子(βNGF)的生物合成。来自标记腺体匀浆的抗βNGF免疫沉淀物的十二烷基硫酸钠凝胶显示出一个单一的主要放射性峰,其与纯化的βNGF迁移一致。加入等量的抗βNGF后,该物质几乎完全沉淀,但加入铁蛋白加抗铁蛋白获得的免疫沉淀物中不存在该物质。标记物质的含胱氨酸胰蛋白酶肽与纯化的βNGF的肽相同。在成年雄性小鼠的下颌下腺中,βNGF的标记约占三氯乙酸可沉淀放射性的0.2%。去势将该值降低到三分之一,而对去势动物进行睾酮处理可使相对βNGF合成恢复到正常或更高水平。在雌性动物的腺体中未检测到βNGF合成。检测了几种组织在培养中合成βNGF的能力。只有下颌下腺将可检测量的放射性掺入βNGF中。通过在体内直接将同位素注射到下颌下腺中也可以获得βNGF的标记。根据βNGF合成与神经元发育和维持的整合情况对结果进行了讨论。