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来自嘧啶生物合成缺陷的中国仓鼠卵巢细胞的多功能蛋白质结构改变。

Alteration in structure of multifunctional protein from Chinese hamster ovary cells defective in pyrimidine biosynthesis.

作者信息

Davidson J N, Patterson D

出版信息

Proc Natl Acad Sci U S A. 1979 Apr;76(4):1731-5. doi: 10.1073/pnas.76.4.1731.

DOI:10.1073/pnas.76.4.1731
PMID:36610
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC383464/
Abstract

A combined genetic, biochemical, and immunological approach has clarified structural relationships involving the first three enzymes of de novo pyrimidine biosynthesis. A procedure involving antibody and protein A-Sepharose was used to isolate the enzymes carbamoyl-phosphate synthase [ATP:carbamate phosphotransferase (dephosphorylating, amido-transferring), EC 2.7.2.9], aspartate transcarbamoyltransferase (carbamoylphosphate:L-aspartate carbamoyltransferase, EC 2.1.3.2), and dihydro-orotase (L-5,6-dihydroorotate amidohydrolase, EC 3.5.2.3) from Chinese hamster ovary cell CHO-K1, the uridine-requiring auxotroph Urd(-)A, and selected Urd(-)A revertants. The enzymes of Urd(-)A and the Urd(-)A revertants were significantly altered in activity, native structure, and molecular weight from those of CHO-K1. The results presented permit the conclusion that (i) these three enzymes reside in a single multifunctional 220,000-dalton polypeptide; (ii) the aspartate transcarbamoyltransferase activity is located on a portion ( approximately 20,000 daltons) at one end of the polypeptide; (iii) this portion may also be required for monomers to aggregate into the multimeric from present in mammalian cells; (iv) the mutations in Urd(-)A and the Urd(-)A revertants lie in the structural gene for this multifunctional protein; and (v) increased sensitivity to proteases could account for the alterations in the structure of these enzymes in the mutants.

摘要

采用遗传学、生物化学和免疫学相结合的方法，阐明了从头嘧啶生物合成过程中前三种酶之间的结构关系。运用一种涉及抗体和蛋白A-琼脂糖凝胶的方法，从中国仓鼠卵巢细胞CHO-K1、尿苷营养缺陷型Urd(-)A以及选定的Urd(-)A回复突变体中分离出氨甲酰磷酸合酶[ATP：氨基甲酸盐磷酸转移酶（去磷酸化，酰胺转移），EC 2.7.2.9]、天冬氨酸转氨甲酰酶（氨甲酰磷酸：L-天冬氨酸氨甲酰转移酶，EC 2.1.3.2）和二氢乳清酸酶（L-5,6-二氢乳清酸酰胺水解酶，EC 3.5.2.3）。Urd(-)A及其回复突变体的这些酶在活性、天然结构和分子量方面与CHO-K1的酶有显著差异。本文给出的结果可以得出以下结论：（i）这三种酶存在于一个单一的220,000道尔顿的多功能多肽中；（ii）天冬氨酸转氨甲酰酶活性位于多肽一端的一部分（约20,000道尔顿）；（iii）该部分可能也是单体聚合成哺乳动物细胞中存在的多聚体形式所必需的；（iv）Urd(-)A及其回复突变体中的突变位于这种多功能蛋白质的结构基因中；（v）对蛋白酶敏感性的增加可以解释突变体中这些酶结构的改变。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/31d4/383464/562bf14f7999/pnas00004-0216-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/31d4/383464/9d7477b2bd6c/pnas00004-0214-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/31d4/383464/574c985f6dc8/pnas00004-0215-a.jpg
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https://cdn.ncbi.nlm.nih.gov/pmc/blobs/31d4/383464/562bf14f7999/pnas00004-0216-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/31d4/383464/9d7477b2bd6c/pnas00004-0214-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/31d4/383464/574c985f6dc8/pnas00004-0215-a.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/31d4/383464/011d88384041/pnas00004-0215-b.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/31d4/383464/97c1783aa02a/pnas00004-0215-c.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/31d4/383464/54e7e94c992a/pnas00004-0215-d.jpg
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本文引用的文献

1
CLONAL GROWTH OF MAMMALIAN CELLS IN A CHEMICALLY DEFINED, SYNTHETIC MEDIUM.哺乳动物细胞在化学成分明确的合成培养基中的克隆生长
Proc Natl Acad Sci U S A. 1965 Feb;53(2):288-93. doi: 10.1073/pnas.53.2.288.
2
Cleavage of structural proteins during the assembly of the head of bacteriophage T4.在噬菌体T4头部组装过程中结构蛋白的切割
Nature. 1970 Aug 15;227(5259):680-5. doi: 10.1038/227680a0.
3
A film detection method for tritium-labelled proteins and nucleic acids in polyacrylamide gels.一种用于检测聚丙烯酰胺凝胶中氚标记蛋白质和核酸的胶片检测方法。
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4
Model involving gene inactivation in the generation of autosomal recessive mutants in mammalian cells in culture.涉及在培养的哺乳动物细胞中产生常染色体隐性突变体时基因失活的模型。
Mol Cell Biol. 1982 Sep;2(9):1126-33. doi: 10.1128/mcb.2.9.1126-1133.1982.
5
Organization of a multifunctional protein in pyrimidine biosynthesis. A domain hypersensitive to proteolysis.嘧啶生物合成中多功能蛋白的组织。对蛋白水解敏感的结构域。
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6
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7
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10
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4
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5
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J Biochem. 1975 Jul;78(1):239-42.
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