Novel recombinant fusion protein analogues of insulin-like growth factor (IGF)-I indicate the relative importance of IGF-binding protein and receptor binding for enhanced biological potency.
Novel recombinant fusion protein analogues of insulin-like growth factor (IGF)-I indicate the relative importance of IGF-binding protein and receptor binding for enhanced biological potency.
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胰岛素样生长因子 (IGF)-I 的新型重组融合蛋白类似物表明 IGF 结合蛋白和受体结合对于增强生物效力的相对重要性。
DOI：
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发表时间：
1992
影响因子：
3.5
通讯作者：
J. Wells
中科院分区：
文献类型：
--
作者：
G. Francis;M. Ross;F. Ballard;S. Milner;C. Senn;K. A. McNeil;J. Wallace;R. King;J. Wells
本文介绍了一种高效表达具有生物活性的胰岛素样生长因子-I（IGF-I）融合肽类似物的大肠杆菌表达系统。这些新的IGF-I融合蛋白类似物具有使它们在IGF-I作用的研究中非常有用的试剂的性质。该类似物包含IGF-1序列和甲硫氨酰猪生长激素（pGH）的前11个氨基酸，并且包括含有真实IGF-1序列的[Met 1]-pGH（1-11）-Val-Asn-IGF-I，以及两种类似物，[Met 1]-pGH（1-11）-Val-Asn-[Gly 3]-IGF-I和[Met 1]-pGH（1-11）-Val-Asn-[Arg 3]-IGF-I，其中人IGF-I序列中的Glu-3分别被Gly或Arg取代。根据存在的IGF-I序列，这三种肽被称为长IGF-I、长[Gly 3]-IGF-I或长[Arg 3]-IGF-I。通过在产生非常高产率的生物活性产物的条件下折叠还原和变性的融合肽序列来辅助纯化的融合肽的生产。引入一个疏水性的N-末端延伸肽似乎有利于正确折叠的IGF-I类似物相比，以前获得的折叠正常长度的IGFs。将IGF-I融合肽的生物活性与真IGF-I和截短类似物des（1-3）IGF-I进行比较。在L 6大鼠成肌细胞中，所有类似物在刺激蛋白质和DNA合成以及抑制蛋白质分解的能力方面都比真正的IGF-I更有效。在H35肝癌细胞中，IGFs通过胰岛素受体起作用，长IGF-I类似物相对于IGF-I保持类似的效力，如在L 6成肌细胞中观察到的。在分泌IGF结合蛋白（IGFBP）到培养基中的细胞系中，生物学效力的顺序为长[Arg 3]-IGF-I-des（1-3）IGF-I大于长[Gly 3]-IGF-I大于长IGF-I大于IGF-I。在鸡胚成纤维细胞中，不向培养基中分泌可检测IGFBPs的细胞系Long [Arg 3]-IGF-I的效力低于IGF-I。通过这些类似物对受体和IGFBP相关性的研究强化了我们之前的发现，即IGF-I的N末端类似物由于IGFBP相互作用程度的变化而显示出更高的生物学效价。
An efficient expression system in Escherichia coli for several biologically active insulin-like growth factor-I (IGF-I) fusion peptide analogues is described. These novel IGF-I fusion protein analogues have properties that make them very useful reagents in the investigation of IGF-I action. The analogues comprise an IGF-I sequence and the first 11 amino acids of methionyl porcine growth hormone (pGH) and include [Met1]-pGH(1-11)-Val-Asn-IGF-I, which contains the authentic IGF-I sequence, and two analogues, [Met1]-pGH(1-11)-Val-Asn-[Gly3]-IGF-I and [Met1]-pGH(1-11)-Val-Asn-[Arg3]-IGF-I, where Glu-3 in the human IGF-I sequence has been replaced by Gly or Arg respectively. The three peptides are referred to as Long IGF-I, Long [Gly3]-IGF-I or Long [Arg3]-IGF-I depending on the IGF-I sequence present. Production of the purified fusion peptides was aided by folding the reduced and denatured fusion peptide sequence under conditions that gave very high yields of biologically active product. Introduction of a hydrophobic N-terminal extension peptide appears to facilitate the correct folding of the IGF-I analogues compared with that obtained previously when folding normal-length IGFs. The biological activities of the IGF-I fusion peptides were compared with authentic IGF-I and the truncated analogue, des(1-3)IGF-I. In L6 rat myoblasts, all the analogues were more potent than authentic IGF-I in their abilities to stimulate protein and DNA synthesis and inhibit protein breakdown. In H35 hepatoma cells, where the IGFs act through the insulin receptor, the Long IGF-I analogues maintained a similar potency relative to IGF-I as was observed in the L6 myoblasts. The order of biological potency in cell lines secreting IGF-binding proteins (IGFBPs) into the medium was Long [Arg3]-IGF-I-des(1-3)IGF-I greater than Long [Gly3]-IGF-I greater than Long IGF-I greater than IGF-I. In chicken embryo fibroblasts, a cell line that does not secrete detectable IGFBPs into the medium, Long [Arg3]-IGF-I, was less potent than IGF-I. Investigation of receptor and IGFBP association by these analogues reinforced our previous findings that N-terminal analogues of IGF-I show increased biological potency due to changes in the degree of their IGFBP interactions.
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II 型胰岛素样生长因子受体不介导 H-35 肝癌细胞中 DNA 合成的增加。
DOI：
--
发表时间：
1984
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Mottola,C;Czech,MP
通讯作者：
Czech,MP
高亲和力胰岛素受体介导大鼠肝癌细胞的生长刺激。
DOI：
--
发表时间：
1982
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
Massagué,J;Blinderman,LA;Czech,MP
通讯作者：
Czech,MP
编码来自大鼠和人类的新型胰岛素样生长因子结合蛋白的 cDNA 的分子克隆。
DOI：
10.1210/mend-4-10-1451
发表时间：
1990
期刊：
Molecular endocrinology (Baltimore, Md.)
影响因子：
--
作者：
Shimasaki,S;Uchiyama,F;Shimonaka,M;Ling,N
通讯作者：
Ling,N
体外肌肉细胞分泌的胰岛素样生长因子结合蛋白类型的鉴定。
DOI：
--
发表时间：
1989
期刊：
The Journal of biological chemistry
影响因子：
--
作者：
McCusker,RH;Camacho-Hübner,C;Clemmons,DR
通讯作者：
Clemmons,DR