Nerve growth factor: biosynthetic products of the mouse salivary glands. Characterization of stable high molecular weight and 32,000-dalton nerve growth factors.
Nerve growth factor: biosynthetic products of the mouse salivary glands. Characterization of stable high molecular weight and 32,000-dalton nerve growth factors.
复制标题
神经生长因子：小鼠唾液腺的生物合成产物。
DOI：
10.1021/bi00367a033
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发表时间：
1986
期刊：
影响因子：
2.9
通讯作者：
Young,M
中科院分区：
文献类型：
--
作者：
Saboori,AM;Young,M
佛罗里达大学医学院，盖恩斯维尔，佛罗里达州 32610 收稿日期：1985 年 12 月 11 日；修订稿于 1986 年 3 月 14 日收到 摘要：神经生长因子 (NGF) 是感觉神经元和交感神经元生长和发育所需的蛋白质。 NGF 在雄性小鼠唾液腺、牛精浆和蛇毒中以高浓度存在。 NGF 在这些来源中的生理意义尚不清楚：它可能是具有可能不同生物功能的高分子量 (HMW) 蛋白质的一部分，并被蛋白酶切割成功能大小。为了分离结构中含有低分子量 (LMW) NGF (2.5 S) 的 HMW 蛋白，在 8 M 尿素或 6 M 盐酸胍 (Gdn-HCl) 存在下对小鼠唾液腺进行均质化，以使蛋白酶变性。该过程表明，LMW NGF 是两种 HMW 蛋白的一部分，这两种蛋白在生物学和免疫学上与小鼠 2.5 S NGF 同源。其中一种 HMW 蛋白（Mr 32 000 NGF）经过纯化，并在 NGF 生物测定中显示出生物活性。此外，该Mr 32 000 NGF被小鼠HMW NGF的y亚基切割成2.5S NGF。还提供证据表明可能存在表观分子量范围为 94 000 至 200 000 且与 7S NGF 的三个亚基（a、0、7）免疫学同源的 HMW 蛋白。该HMW NGF在NGF生物测定中具有生物活性，其活性被0亚基抗体抑制。此外，与小鼠 7S NGF 相比，该 HMWNGF 在 8 M 尿素或 6 M Gdn-HCl 中均不会解离。这两种 HMW NGF 可能在小鼠唾液腺中具有除 7S NGF 亚基所具有的生物学功能之外的生物学功能。神经生长因子是一种调节感觉和交感神经元发育和正常功能的蛋白质，最初由 Levi-Montalcini 和她的同事发现（Levi-Montalcini & Angeletti, 1968）。它以高浓度存在于雄性小鼠唾液腺（Varon 等，1967；Young 等，1978a）、豚鼠前列腺（Harper 等，1979）、牛精浆（Harper 等，1982）和蛇毒（Hogue-Angeletti 等，1976）中。几种培养的细胞系已被证明可以合成 NGF 并将其分泌到培养基中（Bradshaw & Young，1976）。神经生长因子 (NGF) 1 在非神经元来源中的生理意义尚不清楚。这些来源中的 NGF 可能是具有不同生物学功能的 HMW 蛋白的一部分。在小鼠唾液腺中，NGF 是蛋白质的一部分，分子量为 116 000-140 000 (7S NGF)，包含三个不同的亚基：a、0 和 7（Varon 等，1967）。 7S NGF 在 pH 5-8 和高蛋白质浓度（例如 1
College of Medicine, University of Florida, Gainesville, Florida 32610 Received December 11, 1985; Revised Manuscript Received March 14, 1986 abstract: Nerve growth factor (NGF) is a protein required for the growth and development of sensory and sympathetic neurons. The NGF is present in high concentrations in male mouse salivary glands, bovine seminal plasma, and snake venom. The physiological significance of NGF in these sources is not known: it might be a part of a high molecular weight (HMW) protein with possibly different biological function and be cleaved tothe functional size by proteases. In an attempt to isolate a HMW protein containing as part of itsstructure the low molecular weight (LMW) NGF (2.5 S), mouse salivary glands were homogenized in the presence of either 8 M urea or 6 M guanidine hydrochloride (Gdn-HCl) in order to denature proteases. This procedure revealed that the LMW NGF is a part of two HMW proteins that are biologically and immunologically homologous to the mouse 2.5 S NGF. One of these HMW proteins (Mr 32 000 NGF) was purified and shown to be biologically active inthe NGF bioassay. Furthermore, this Mr 32 000 NGF was cleaved by the y subunit of mouse HMW NGF to the 2.5 S NGF. Evidence is also presented that there may be a HMW protein (s) with apparent molecular weights ranging from 94 000 to 200 000 and immunologically homologousto the three subunits (a, 0, 7) of 7S NGF. This HMW NGF is biologically active in the NGF bioassay, and its activity is inhibited by antibody to the 0 subunit. Furthermore, in contrast to mouse 7S NGF, this HMWNGF does notdissociate in either 8 M urea or 6 M Gdn-HCl. These two HMW NGF may have biological functions in the mouse salivary glands other than those that have been attributed to the subunits of 7S NGF. e nerve growth factor, a protein that regulates the de-velopment and normal function of sensory and sympathetic neurons, was originally discovered by Levi-Montalcini and her colleagues (Levi-Montalcini & Angeletti, 1968). It is present at high concentrations in male mouse salivary glands (Varon et al., 1967; Young et al., 1978a), guinea pig prostate glands (Harper et al., 1979), bovine seminal plasma (Harper et al., 1982), and snake venom(Hogue-Angeletti et al., 1976). Several cultured cell lines have been shown to synthesize and secrete NGF into the culture medium (Bradshaw & Young, 1976).The physiological significance of nerve growth factor (NGF) 1 in nonneuronal sources is not known. The NGF in these sources might be a part of a HMW protein with different biological function (s). In the mouse salivary glands, the NGF is a part of a protein, molecular weight of 116 000-140 000 (7S NGF), that contains three different subunits, a, 0, and 7 (Varon et al., 1967). The 7S NGF is stable at pH 5-8 and at a high protein concentration (eg, 1