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ISOLATION AND RECONSTITUTION OF SUGAR TRANSPORT CARRIER

糖运输载体的隔离和重建

基本信息

批准号：
3150863
负责人：
CHAN Y. JUNG
金额：
$ 8.12万
依托单位：
STATE UNIVERSITY OF NEW YORK AT BUFFALO
依托单位国家：
美国
项目类别：
财政年份：
1978
资助国家：
美国
起止时间：
1978-07-01 至 1987-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/3150863
关键词：
Ehrlich's tumor affinity chromatography affinity labeling analytical method blood /lymphatic pharmacology chemical structure function crosslink cytochalasins erythrocyte membrane glucose transport human tissue liposomes membrane permeability membrane reconstitution /synthesis photochemistry physical separation radiotracer

项目摘要

This research program is a continuous effort to isolate and reconstitute the structural unit of glucose transport carrier function of human erythrocytes, and to characterize its operation at the molecular level. To this end, we propose the following specific points to be studied: 1) To characterize the "180,000 dalton-membrane protein," which we have differentially labeled by fluorodinitrobenzene and tentatively proposed to be glucose carrier. This would include an isolation on a preparative scale; chemical and physical characterization; and reconstitution of the carrier activity in in vitro membrane structures from the protein. 2) To isolate and characterize the "glucose-sensitive, cytochalasin B binding protein," which we have also tentatively proposed to be the carrier, by means of covalent labeling or affinity chromatography. This would involve synthesis of protein-reactive cytochalasin B derivatives as covalent reagents; structure-activity relationship studies on cytochalasin B and the derivatives, both in terms of the glucose-carrier inhibition and of the glucose-sensitive binding; isolation and identification by covalent labeling and/or affinity chromatography; and characterization of the binding protein in terms of the glucose carrier activity in a reconstituted in vitro system. 3) To study the mode of information-transduction from insulin-receptor to glucose carrier, in native and in model membrane systems. This would include isolation of the insulin receptors; attempts at isolation of "180,000 dalton-protein" and "glucose-sensitive, cytochalasin B binding protein," both from fat cell membranes; and subsequent characterization of interaction of these proteins with insulin receptors.

这项研究计划是一项持续的努力，以分离和重组人红细胞葡萄糖转运载体功能的结构单元，和来描述它在分子水平上的运作。为此，我们建议具体有以下几点需要研究：1）对“18万道尔顿膜蛋白，”我们已经区别标记，氟代二硝基苯，并初步提出作为葡萄糖载体。这将包括制备规模分离;化学和物理分离表征;以及在体外重建载体活性蛋白质的膜结构。 2)为了分离和表征 “葡萄糖敏感的细胞松弛素B结合蛋白”，我们也有暂时提出作为载体，通过共价标记或亲和层析这将涉及合成蛋白质反应作为共价试剂的细胞松弛素B衍生物;构效关系细胞松弛素B及其衍生物的研究，无论是在葡萄糖载体抑制和葡萄糖敏感性结合;分离和通过共价标记和/或亲和色谱法鉴定;和结合蛋白在葡萄糖载体活性方面的表征在体外重建的系统中。 3)研究模式从胰岛素受体到葡萄糖载体的信息转导，在天然和在模型膜系统中。这将包括分离胰岛素受体;试图分离“180，000道尔顿蛋白质”， “葡萄糖敏感性细胞松弛素B结合蛋白”，均来自脂肪细胞膜;以及随后表征这些蛋白质与胰岛素受体

项目成果

期刊论文数量（40）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Structural basis of human erythrocyte glucose transporter function in reconstituted vesicles.

重组囊泡中人红细胞葡萄糖转运蛋白功能的结构基础。

DOI：
发表时间：
1986
期刊：
The Journal of biological chemistry
影响因子：
0
作者：
Chin,JJ;Jung,EK;Jung,CY
通讯作者：
Jung,CY

Hypertonic cryohemolysis and the cytoskeletal system.

高渗冷冻溶血和细胞骨架系统。

DOI：
10.1016/0005-2736(81)90038-9
发表时间：
1981
期刊：
Biochimica et biophysica acta
影响因子：
0
作者：
Green,FA;Jung,CY;Cuppoletti,J;Owens,N
通讯作者：
Owens,N

Target size of 5'-nucleotidase in smooth muscle.

平滑肌中 5-核苷酸酶的目标大小。

DOI：
10.1093/oxfordjournals.jbchem.a135312
发表时间：
1985
期刊：
Journal of biochemistry
影响因子：
2.7
作者：
Grover,AK;Agrawal,DK;Ahmad,S;Daniel,EE;Kwan,CY;Oakes,PJ;Sipos,SN;Berenski,C;Jung,C
通讯作者：
Jung,C

Structural basis of human erythrocyte glucose transporter function in proteoliposome vesicles: circular dichroism measurements.

蛋白脂质体囊泡中人红细胞葡萄糖转运蛋白功能的结构基础：圆二色性测量。

DOI：
10.1073/pnas.84.12.4113
发表时间：
1987
期刊：
Proceedings of the National Academy of Sciences of the United States of America
影响因子：
11.1
作者：
Chin,JJ;Jung,EK;Chen,V;Jung,CY
通讯作者：
Jung,CY

Molecular properties of the slow inward calcium channel. Molecular weight determinations by radiation inactivation and covalent affinity labeling.

慢向内钙通道的分子特性。