Assignment of the human gene for liver-type 6-phosphofructokinase isozyme (PFKL) to chromosome 21 by using somatic cell hybrids and monoclonal anti-L antibody.
Assignment of the human gene for liver-type 6-phosphofructokinase isozyme (PFKL) to chromosome 21 by using somatic cell hybrids and monoclonal anti-L antibody.
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使用体细胞杂交体和单克隆抗 L 抗体将人类肝型 6-磷酸果糖激酶同工酶 (PFKL) 基因分配至 21 号染色体。
DOI：
10.1073/pnas.78.6.3738
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发表时间：
1981
影响因子：
11.1
通讯作者：
Francke,U
中科院分区：
文献类型：
--
作者：
Vora,S;Francke,U
人6-磷酸果糖激酶（PFK； ATP: d -果糖-6-磷酸1-磷酸转移酶，EC 2.7.1.11）受编码肌肉(M)、肝脏(L)和血小板(P)亚基的结构位点控制，这些亚基在不同组织中表达不同；人类二倍体成纤维细胞和白细胞表达这三种基因。这些亚基的随机四聚化产生各种同工酶，它们可以通过离子交换色谱法或亚基特异性单克隆抗体相互区分。我们检测了中国仓鼠细胞与人二倍体成纤维细胞或白细胞之间建立的17个体细胞杂交，以表达人PFK的l型亚基。由于电泳无法区分中国仓鼠PFKs和人PFKs，我们使用了一种抗人l -亚基特异性单克隆抗体，该抗体与中国仓鼠PFKs不发生反应。以含蛋白A葡萄球菌为免疫吸附剂，采用酶免疫沉淀法检测杂交体中人L亚基的表达。通过染色体和同工酶标记分析，17个杂种中有12个表达人的L亚基，保留了21号染色体，而5个不表达人的PFKL，缺乏21号染色体。7例21三体患者的平均红细胞PFK升高（为正常人的147%）。三体红细胞中L亚基的特异性增加在色谱上明显表现为L4种显著增加（50%，正常10%），免疫上表现为抗m单克隆抗体沉淀减少（50%，正常80%）。我们从这些数据中得出结论，PFKL位于21号染色体上，并且先前注意到的21三体患者红细胞PFK活性的升高是由于基因剂量效应。
Human 6-phosphofructokinase (PFK; ATP:D-fructose-6-phosphate 1-phosphotransferase, EC 2.7.1.11) is under the control of structural loci that code for muscle (M), liver (L), and platelet (P) subunits, which are variably expressed in different tissues; human diploid fibroblasts and leukocytes express all three genes. Random tetramerization of these subunits produces various isozymes, which can be distinguished from one another by ion exchange chromatography or by subunit-specific monoclonal antibodies. We have examined 17 somatic cell hybrids established between Chinese hamster cells and human diploid fibroblasts or leukocytes for the expression of L-type subunits of human PFK. As electrophoresis does not distinguish between Chinese hamster PFKs and human PFKs, we used an anti-human L-subunit-specific monoclonal antibody, which does not react with chinese hamster PFKs. The expression of human L subunits in the hybrids was detected by the enzyme-immunoprecipitation technique using staphylococci bearing protein A as an immunoadsorbent. Twelve out of 17 hybrids expressed human L subunits and retained chromosome 21, as determined by chromosome and isozyme marker analysis, whereas 5 did not express human PFKL and lacked chromosome 21. The mean erythrocyte PFK of seven individuals with trisomy 21 was found to be elevated (147% of normal). A specific increase in L subunits in trisomic erythrocytes was evident chromatographically by a striking increase in L4 species (50%; normal 10%) and immunologically by decreased precipitation with anti-M monoclonal antibody (50%; normal 80%). We conclude from these data that PFKL is located on chromosome 21 and that the previously noted elevation of erythrocyte PFK activity in individuals with trisomy 21 is due to a gene-dosage effect.