Characterization of cDNA clones for human myeloperoxidase: predicted amino acid sequence and evidence for multiple mRNA species.
Characterization of cDNA clones for human myeloperoxidase: predicted amino acid sequence and evidence for multiple mRNA species.
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人髓过氧化物酶 cDNA 克隆的表征：预测的氨基酸序列和多种 mRNA 种类的证据。
DOI：
10.1093/nar/15.5.2013
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发表时间：
1987
影响因子：
14.9
通讯作者：
Rovera,G
中科院分区：
文献类型：
--
作者：
Johnson,KR;Nauseef,WM;Care,A;Wheelock,MJ;Shane,S;Hudson,S;Koeffler,HP;Selsted,M;Miller,C;Rovera,G
髓过氧化物酶是多形核白细胞杀微生物网络的一个组成部分。该酶是由两个重亚基和两个轻亚基组成的四聚体。很大一部分人表现出髓过氧化物酶产生的遗传缺陷。作为更详细地分析这些缺陷的第一步，我们从HL-60人早幼粒细胞白血病细胞系的表达库中分离了髓过氧化物酶的cDNA克隆。两个重叠的质粒用髓过氧化物酶抗血清检测1）在含pMP 02的细菌中表达的70 kDa蛋白和2）通过使用pMP 062和HL-60 RNA的杂交选择和翻译产生的75 kDa多肽。通过将预测的氨基酸序列与重亚基和轻亚基的氨基末端序列进行匹配来进行克隆的正式鉴定。这两个亚基由一个mRNA按以下顺序编码：前体-原序列-轻亚基-重亚基，预测的初级翻译产物的分子量为1 s 83.7 kDa。北方印迹揭示了两种大小的杂交RNA（大约3.0-3.3和3.5-4.0个内切酶），其表达限于粒细胞谱系的细胞，并且与分化期间观察到的酶活性的变化平行。
Myeloperoxidase is a component of the microbicidal network of polymorphonuclear leukocytes. The enzyme is a tetramer consisting of two heavy and two light subunits. A large proportion of humans demonstrate genetic deficiences in the production of myeloperoxidase. As a first step in analyzing these deficiencies 1n more detail, we have isolated cDNA clones for myeloperoxidase from an expression library of the HL-60 human promyelocytic leukemia cell line.Two overlapping plasmids(pMP02 and pMP062) were identified as myeloperoxidase cDNA clones based on the detection with myeloperoxidase antiserum of 1)70 kDa protein expressed 1n pMP02-conta1n1ng bacteria and 2) a 75 kDa polypeptide produced by hybridization selection and translation using pMP062 and HL-60 RNA. Formal identification of the clones was made by matching the predicted amino add sequences with the amino terminal sequences of the heavy and light sub-units. Both subunits are encoded by one mRNA in the following order: pre-pro-sequences -- light subunit -- heavy subunit.The molecular weight of the predicted primary translation product 1s 83.7 kDa. Northern blots reveal two size classes of hybridizing RNAs(approximately 3.0–3.3 and 3.5–4.0 kilobases) whose expression is restricted to cells of the granulocytic lineage and parallels the changes in enzymatic activity observed during differentiation.