125I-glycoconjugate labels for identifying sites of protein catabolism in vivo: effect of structure and chemistry of coupling to protein on label entrapment in cells after protein degradation.
125I-glycoconjugate labels for identifying sites of protein catabolism in vivo: effect of structure and chemistry of coupling to protein on label entrapment in cells after protein degradation.
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用于识别体内蛋白质分解代谢位点的 125I-糖缀合物标签：蛋白质偶联的结构和化学对蛋白质降解后细胞中标签捕获的影响。
DOI：
10.1016/0003-9861(85)90071-2
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发表时间：
1985
影响因子：
3.9
通讯作者：
Thorpe,SR
中科院分区：
文献类型：
--
作者：
Strobel,JL;Baynes,JW;Thorpe,SR
残留放射性标记被设计成在载体蛋白降解后仍被困在细胞内，并已用于识别血浆蛋白分解代谢的组织和细胞部位。在这项研究中，我们描述了一系列125I标记的糖偶联物的简便合成和纯化，并评价了它们在模型载体蛋白asialofetuin降解后在肝脏中的滞留效率。用NaBH3CN还原芳香胺，以65%-90%的产率合成了糖结合物。产物经一次离子交换层析纯化，然后用125I标记。所制备的衍生物是单取代和双取代乳糖醇、纤维二醇和葡糖醇-[125I]酪胺和乳糖醇-[125I]酪氨酸。125I-糖偶联物用三聚氯氰偶联到去唾液酸上，对于含乳糖的标记物，先用半乳糖氧化酶处理，然后用NaBH3CN还原胺化。通过这两种方法粘贴标记都不会影响积雪草素从大鼠循环中的正常快速清除，也不会影响其在肝脏溶酶体中的摄取和降解。通过跟踪全身放射性损失的动力学，测量~(125)I标记的降解产物从细胞中的渗漏。我们观察到，来自较大的双取代糖共轭化合物的降解产物比来自较小的和单取代的衍生物的降解产物被更有效地保留，并且通过还原胺化与蛋白质偶联的糖共轭化合物比通过三聚氯氰偶联的糖共轭化合物更有效地保留在体内。总体而言，缩乳醇-[125I]酪胺通过还原氨化偶联到蛋白质上，在肝脏中的包封率最高。
Residualizing radioactive labels are designed to remain entrapped within cells following degradation of a carrier protein, and have been used for identification of the tissue and cellular sites of plasma protein catabolism. In this study we describe a convenient synthesis and purification of a series of125I-labeled glycoconjugates, and an evaluation of their efficiency of retention in liver following degradation of a model carrier protein, asialofetuin. Glycoconjugates were prepared in 65–90% yield by reductive animation of reducing sugars with aromatic amines using NaBH3CN. The products were purified in a single ion-exchange chromatographic step, and then labeled with125I. The derivatives prepared were mono- and disubstituted lactitol-, cellobiitol- and glucitol-[125I]tyramine and lactitol-[125I]tyrosine.125I-Glycoconjugates were coupled to asialofetuin using either cyanuric chloride or, for lactose-containing labels, by treatment with galactose oxidase followed by reductive amination with NaBH3CN. Attachment of labels by either procedure did not affect the normal rapid clearance of asialofetuin from the rat circulation nor its uptake and degradation in liver lysosomes. Leakage of125I-labeled degradation products from cells was measured by following the kinetics of loss of whole-body radioactivity. We observed that degradation products from larger, disubstituted glycoconjugates were retained more efficiently than those from smaller and monosubstituted derivatives, and that glycoconjugates coupled to protein via reductive amination were retained in the body more efficiently than those coupled by cyanuric chloride. Overall, dilactitol-[125I]tyramine coupled to protein by reductive amination was entrapped most efficiently in liver.
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放射性碘标记蛋白质降解产物的测量。
影响因子：
2.9
作者：
G. Chisolm;C. Sila;S. Hmiel
通讯作者：
S. Hmiel
一种放射性碘标记的细胞内捕获配体，用于确定体内血浆蛋白降解的位点。
DOI：
10.1042/bj2120791
发表时间：
1983
期刊：
The Biochemical journal
影响因子：
--
作者：
Pittman,RC;Carew,TE;Glass,CK;Green,SR;TaylorJr,CA;Attie,AD
通讯作者：
Attie,AD
DOI：
10.1139/o67-217
发表时间：
1967-01-01
期刊：
CANADIAN JOURNAL OF BIOCHEMISTRY
影响因子：
--
作者：
GREGORIA.G;SOURKES, TL
通讯作者：
SOURKES, TL
注射蛋白质的选择性降解主要发生在胞质溶胶中，而不是溶酶体中
影响因子：
64.5
作者：
S. Bigelow;R. Hough;M. Rechsteiner
通讯作者：
M. Rechsteiner
水溶性半乳甘露聚糖化学成分与旋光度的相关性
影响因子：
3.1
作者：
C. Leschziner;A. Cerezo
通讯作者：
A. Cerezo