Saito Y, Tsubuki S, Ito H, Kawashima S
Department of Enzyme Biochemistry, Tokyo Metropolitan Institute of Gerontology, Japan.
Biochem Biophys Res Commun. 1992 Apr 15;184(1):419-26. doi: 10.1016/0006-291x(92)91210-h.
A tripeptide protease inhibitor, benzyloxycarbonyl-Leu-Leu-Leu-aldehyde (ZLLLal), induces the outgrowth of one or two long neurites from PC12 cells. Since this neurite outgrowth is different from that induced by nerve growth factor (NGF) in some aspects, the existence of a molecule that regulates neurite formation in PC12 cells was expected. To identify a target molecule, Leu-Leu-Leu-aldehyde (LLLal) was immobilized as a ligand for affinity chromatography. Proteins of 33-kDa, 35-kDa, and 180-kDa from the membrane and cytoplasmic fractions of PC12 cells bound specifically to the affinity column. ZLLL-COOH has no ability to induce neurite outgrowth, and the 33-kDa, 35-kDa, and 180-kDa proteins do not bind to an LLL-COOH coupled affinity column. By using the LLLal-affinity column, the 33-kDa/35-kDa proteins were found to be converted to 36-kDa/38-kDa proteins during brain development in rats. These results suggest that LLLal-binding proteins are involved in neuronal differentiation.
一种三肽蛋白酶抑制剂，苄氧羰基 - 亮氨酸 - 亮氨酸 - 亮氨酸 - 醛（ZLLLal）可诱导PC12细胞长出一到两条长神经突。由于这种神经突生长在某些方面与神经生长因子（NGF）诱导的不同，因此预期存在一种调节PC12细胞中神经突形成的分子。为了鉴定靶分子，将亮氨酸 - 亮氨酸 - 亮氨酸 - 醛（LLLal）固定为亲和色谱的配体。来自PC12细胞膜和细胞质部分的33 kDa、35 kDa和180 kDa蛋白质与亲和柱特异性结合。ZLLL - COOH没有诱导神经突生长的能力，并且33 kDa、35 kDa和180 kDa蛋白质不与LLL - COOH偶联的亲和柱结合。通过使用LLLal - 亲和柱，发现大鼠脑发育过程中33 kDa / 35 kDa蛋白质转化为36 kDa / 38 kDa蛋白质。这些结果表明LLLal结合蛋白参与神经元分化。
