Rat serum factors inhibiting the G1-S transition in hepatocytes. II. Properties of the low molecular weight factor.

Cell and tissue kinetics Pub Date : 1983-07-01
E Le Rumeur, J J Winchenne, G A Boffa, C Nadal
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Abstract

The properties were investigated of low molecular weight factors acting on the G1-S transition of baby rat hepatocytes. These factors were produced by incubating adult rat serum with trypsin or a 100,000 g liver microsomal fraction, and isolated by ultrafiltration. Enzyme degradation assays indicated that the active compound was in both cases a glycopeptide sensitive to pronase and papain and to the combination of neuraminidase and beta galactosidase. No loss of hepatocyte G1-S inhibitory activity was observed after heat treatment at pH 7.0. G50 gel filtration showed that both the G1-S inhibitory factors were collected in the last fractions eluted. The elution volume was slightly larger for the trypsin than for the microsomal-induced factor, suggesting a small difference between their molecular weight. These factors are believed to be glycopeptides with a molecular weight around 1400. They might be composed of a 3-sugar polysaccharide chain with a galactose preterminal and a neuraminic acid terminal, linked to a polypeptide chain of 6 to 8 amino acids.

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抑制肝细胞G1-S转化的血清因子。2低分子量因子的性质。
研究了低分子量因子对幼鼠肝细胞G1-S转变的影响。这些因子是通过用胰蛋白酶或100,000 g肝微粒体片段孵育成年大鼠血清产生的,并通过超滤分离。酶降解实验表明,活性化合物在两种情况下都是对蛋白酶和木瓜蛋白酶敏感的糖肽,对神经氨酸酶和半乳糖苷酶的组合敏感。pH 7.0热处理后肝细胞G1-S抑制活性未见下降。G50凝胶过滤结果显示,洗脱后的组分中均有G1-S抑制因子。胰蛋白酶的洗脱体积略大于微粒体诱导因子的洗脱体积,这表明它们的分子量之间存在微小差异。这些因素被认为是分子量在1400左右的糖肽。它们可能由具有半乳糖前端和神经氨酸末端的3糖多糖链组成,与6至8个氨基酸的多肽链相连。
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