Cloning, expression and chromosomal localization of a human testis 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase gene
Cloning, expression and chromosomal localization of a human testis 6-phosphofructo-2-kinase/fructose-2,6-bisphosphatase gene
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DOI：
10.1016/s0378-1119(99)00037-2
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发表时间：
1999-03-18
期刊：
影响因子：
3.5
通讯作者：
Bartrons, R
中科院分区：
文献类型：
--
作者：
Manzano, A;Pérez, JX;Bartrons, R
6-磷酸果糖-2-激酶/果糖2，6-二磷酸酶（PFK-2/FBPase-2）是一种双功能酶，负责合成和分解糖酵解的关键代谢产物Fru-2，6-P-2。几个基因编码不同的PFK-2/FBPase-2同工酶，其组织分布和酶调节不同。在本文中，我们提出了从人睾丸cDNA文库中分离的cDNA编码PFK-2/FBPase-2同工酶。测序数据显示编码469个氨基酸的蛋白质的1407个核苷酸的开放阅读框。这与大鼠睾丸PFK-2/FBPase-2具有54 kDa的计算分子量和97%的相似性，其中参与催化机制的氨基酸残基完全保守。荧光原位杂交（FISH）将睾丸PFK-2/FBPase-2基因（PFKFB-4）定位于人3号染色体p21-p22带。北方印迹分析表明，在不同的大鼠组织中存在的2.4 kb的mRNA特异性表达在睾丸。在哺乳动物COS-1细胞中，人睾丸cDNA驱动分子量为55 kDa的同工酶的表达。该同工酶具有明显的PFK-2活性。综上所述，这些结果提供了一个新的PFK-2/FBPase-2基因编码的人睾丸同工酶的证据。(C)1999 Elsevier Science B. V.保留所有权利。
6-Phosphofructo-2-kinase/fructose 2,6-bisphosphatase (PFK-2/FBPase-2) is a bifunctional enzyme responsible for the synthesis and breakdown of Fru-2,6-P-2, a key metabolite in the regulation of glycolysis. Several genes encode distinct PFK-2/FBPase-2 isozymes that differ in their tissue distribution and enzyme regulation. In this paper, we present the isolation of a cDNA from a human testis cDNA library that encodes a PFK-2/FBPase-2 isozyme. Sequencing data show an open reading frame of 1407 nucleotides that codifies for a protein of 469 amino acids. This has a calculated molecular weight of 54 kDa and 97% similarity with rat testis PFK-2/FBPase-2, with complete conservation of the amino acid residues involved in the catalytic mechanism. Fluorescence in-situ hybridization (FISH) localized testis PFK-2/FBPase-2 gene (PFKFB4) in human chromosome 3 at bands p21-p22. A Northern blot analysis of different rat tissues showed the presence of a 2.4-kb mRNA expressed specifically in testis. In mammalian COS-1 cells, the human testis cDNA drives expression of an isozyme with a molecular weight of 55 kDa. This isozyme shows clear PFK-2 activity. Taken together, these results provide evidence for a new PFK-2/FBPase-2 gene coding for a human testis isozyme. (C) 1999 Elsevier Science B.V. All rights reserved.