"STRUCTURE-FUNCTION STUDIES OF DIHYDROFOLATE REDUCTASE"
"STRUCTURE-FUNCTION STUDIES OF DIHYDROFOLATE REDUCTASE"
批准号:
3181953
负责人:
JAMES H. FREISHEIM
金额:
$17.14万
依托单位国家:
美国
项目类别:
财政年份:
1985
资助国家:
美国
项目状态:
已结题
起止时间:
1985-05-01 至 1988-04-30
中文摘要
本项目关注的是结构与
二氢叶酸还原酶的功能(二氢叶酸+NADPH + H +-)
四氢叶酸+NADP+)。 利用蛋白质的一些技术
化学,要研究的问题包括:(a)获得蛋白质
甲氨蝶呤(MTX)不敏感酶的序列信息,
MTX抗性L5178 Y细胞和来自步行者256癌肉瘤细胞
主要通过衍生肽的HPLC图谱和氨基酸分析;(B)
蛋白质的特定氨基酸残基的化学修饰,
测量这些改性对基材的影响,
辅因子和抑制剂结合以及对酶活性的影响;(c)平衡
使用还原酶进行底物、辅因子和抑制剂的结合研究,
人、L5178 Y和步行者256细胞,并使用荧光、圆形
二色性和平衡透析技术;(d)MTX的评价
作为二氢叶酸还原酶的亲和和光亲和标记的类似物;
(e)选择的取代的喹唑啉、三嗪和
作为酶抑制剂的蝶啶及其转运的测定
MTX敏感和耐药细胞的特征。
英文摘要
This project is concerned with the relation of the structure to the
function of dihydrofolate reductase (dihydrofolate+ NADPH + H+ -
tetrahydrofolate + NADP+). Using a number of techniques of protein
chemistry, the problems to be investigated include: (a) obtaining protein
sequence information on a methotrexate (MTX)-insensitive enzyme from
MTX-resistant L5178Y cells and from Walker 256 carcinosarcoma cells
primarily by HPLC-mapping of derived peptides and amino acid analysis; (b)
chemical modification of specific amino acid residues of the protein and
measurements of the effect of these modifications on the substrate,
cofactor and inhibitor binding and on enzymic activity; (c) equilibrium
binding studies of substrate, cofactor and inhibitors using reductases from
human, L5178Y and Walker 256 cells and employing fluorescence, circular
dichroism and equilibrium dialysis techniques; (d) evaluation of MTX
analogues as affinity and photoaffinity labels of dihydrofolate reductases;
(e) examination of selected, substituted quinazolines, triazines and
pteridines as enzyme inhibitors and a determination of their transport
characteristics in MTX-sensitive and -resistant cells.
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"STRUCTURE-FUNCTION STUDIES OF DIHYDROFOLATE REDUCTASE"
-
批准号:3181956
-
项目类别:
-
资助金额:$18.57万
-
财政年份:1985
-
负责人:JAMES H. FREISHEIM
-
依托单位:
STRUCTURE-FUNCTION STUDIES OF DIHYDROFOLATE REDUCTASE
-
批准号:3181954
-
项目类别:
-
资助金额:$24.43万
-
财政年份:1985
-
负责人:JAMES H. FREISHEIM
-
依托单位:
STRUCTURE-FUNCTION STUDIES OF DIHYDROFOLATE REDUCTASE
-
批准号:3181960
-
项目类别:
-
资助金额:$23.88万
-
财政年份:1985
-
负责人:JAMES H. FREISHEIM
-
依托单位:
"STRUCTURE-FUNCTION STUDIES OF DIHYDROFOLATE REDUCTASE"
-
批准号:3181955
-
项目类别:
-
资助金额:$17.81万
-
财政年份:1985
-
负责人:JAMES H. FREISHEIM
-
依托单位:
STRUCTURE-FUNCTION STUDIES OF DIHYDROFOLATE REDUCTASE
-
批准号:3181957
-
项目类别:
-
资助金额:$22.18万
-
财政年份:1985
-
负责人:JAMES H. FREISHEIM
-
依托单位:
STRUCTURE-FUNCTION STUDIES OF DIHYDROFOLATE REDUCTASE
-
批准号:3181959
-
项目类别:
-
资助金额:$22.96万
-
财政年份:1985
-
负责人:JAMES H. FREISHEIM
-
依托单位:
STRUCTURE-FUNCTION STUDIES OF DIHYDROFOLATE REDUCTASE
-
批准号:3181958
-
项目类别:
-
资助金额:$22.07万
-
财政年份:1985
-
负责人:JAMES H. FREISHEIM
-
依托单位:
海外基金