Biosynthesis of the low molecular weight carrier protein for insulin-like growth factors in rat liver and fibroblasts.
Biosynthesis of the low molecular weight carrier protein for insulin-like growth factors in rat liver and fibroblasts.
复制标题
大鼠肝脏和成纤维细胞中胰岛素样生长因子的低分子量载体蛋白的生物合成。
作者：
J. A. Romanus;Y. Yang;S. Nissley;M. Rechler
通过完整细胞的生物合成标记和提取RNA的无细胞翻译，研究了胰岛素样生长因子（IGFs）的低分子量（Mr）载体蛋白在BRL-3A大鼠肝细胞系、大鼠胚胎成纤维细胞（REFs）和胎鼠肝中的生物合成。[35 S]半胱氨酸标记的载体蛋白前体使用从BRL-3A细胞产生的约33，000 Mr载体蛋白的抗体进行免疫沉淀，该抗体识别存在于胎儿和新生大鼠血清中的IGF载体蛋白，但不识别成年大鼠血清中的IGF载体蛋白。IGF载体蛋白在由来自BRL-3A细胞或REF的RNA指导的网织红细胞裂解物翻译系统中合成为35，000 Mr前体。补充翻译孵育与微粒体膜的大小减少前体的33,000先生，大概是通过去除信号肽。在完整BRL-3A细胞或REF的连续标记或脉冲追逐实验中，33，000 Mr蛋白在10分钟内被细胞内标记，40分钟后出现在培养基中，并在培养基中持续24小时，大小没有变化。在BRL-3A细胞中用[3 H]亮氨酸、[3 H]苯丙氨酸、[3 H]精氨酸或[35 S]半胱氨酸生物合成标记细胞内载体蛋白并纯化，并测定其NH 2-末端氨基酸序列。34个残基中的11个被鉴定，并且对应于从条件培养基纯化的成熟未标记的载体蛋白的残基，这表明在去除信号肽后，载体蛋白在其NH 2-末端不经历可检测的进一步加工。这些结果表明，虽然他们协调调节，IGF-II和胎儿IGF载体蛋白作为单独的蛋白质合成。最后，从胎儿，但不是成年大鼠肝脏提取的RNA指导合成的35,000先生载体蛋白前体，这表明载体蛋白的发育调节可能发生在RNA丰度的水平。
Biosynthesis of the low mol wt (Mr) carrier protein for insulin-like growth factors (IGFs) was studied in the BRL-3A rat liver cell line, rat embryo fibroblasts (REFs), and fetal rat liver by biosynthetic labeling of intact cells and cell-free translation of extracted RNA. [35S]Cysteine-labeled carrier protein precursors were immunoprecipitated using antibodies raised to the approximately 33,000 Mr carrier protein from BRL-3A cells that recognize the IGF carrier protein present in fetal and neonatal rat serum, but not in adult rat serum. The IGF carrier protein is synthesized as a 35,000 Mr precursor in a reticulocyte lysate translation system directed by RNA from BRL-3A cells or REFs. Supplementation of the translation incubation with microsomal membranes decreases the size of the precursor to 33,000 Mr, presumably by removal of a signal peptide. In continuous labeling or pulse-chase experiments of intact BRL-3A cells or REFs, the 33,000 Mr protein is labeled within 10 min intracellularly, appears in the medium after 40 min, and persists in the medium for 24 h without a change in size. The intracellular carrier protein was biosynthetically labeled in BRL-3A cells with [3H]leucine, [3H]phenylalanine, [3H]arginine, or [35S]cysteine and purified, and its NH2-terminal amino acid sequence was determined. Eleven of 34 residues were identified and correspond to those of mature unlabeled carrier protein purified from conditioned medium, indicating that after removal of the signal peptide, the carrier protein undergoes no detectable further processing at its NH2-terminus. These results establish that although they are regulated coordinately, IGF-II and the fetal IGF carrier protein are synthesized as separate proteins. Finally, RNA extracted from fetal, but not adult, rat liver directs the synthesis of the 35,000 Mr carrier protein precursor, suggesting that the developmental regulation of the carrier protein may occur at the level of RNA abundance.
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DOI：
--
发表时间：
1985
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Massagué,J;Kelly,B;Mottola,C
通讯作者：
Mottola,C
DOI：
10.1016/s0021-9258(18)67484-8
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1986-08
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Charles T. Roberts;A. L. Brown;D. Graham;Steven Seelig;Susan A. Berry;K. Gabbay;M. Rechler
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Charles T. Roberts;A. L. Brown;D. Graham;Steven Seelig;Susan A. Berry;K. Gabbay;M. Rechler
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DOI：
10.1172/jci112343
发表时间：
1986
期刊：
The Journal of clinical investigation
影响因子：
--
作者：
DeVroede,MA;Tseng,LY;Katsoyannis,PG;Nissley,SP;Rechler,MM
通讯作者：
Rechler,MM
DOI：
10.1089/dna.1983.2.329
发表时间：
1983-01-01
期刊：
DNA-A JOURNAL OF MOLECULAR & CELLULAR BIOLOGY
影响因子：
--
作者：
CATHALA, G;SAVOURET, JF;BAXTER, JD
通讯作者：
BAXTER, JD
大鼠肝 BRL-3A 细胞分泌的胰岛素样生长因子结合蛋白的纯化和氨基末端序列。
DOI：
--
发表时间：
1986
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Mottola,C;MacDonald,RG;Brackett,JL;Mole,JE;Anderson,JK;Czech,MP
通讯作者：
Czech,MP