Mouse N-acetylgalactosamine 4-sulfotransferases-1 and -2. Molecular cloning, expression, chromosomal mapping and detection of their activity with GalNAcbeta1-4GlcNAcbeta1-octyl.
Mouse N-acetylgalactosamine 4-sulfotransferases-1 and -2. Molecular cloning, expression, chromosomal mapping and detection of their activity with GalNAcbeta1-4GlcNAcbeta1-octyl.
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小鼠 N-乙酰半乳糖胺 4-磺基转移酶-1 和 -2。
DOI：
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发表时间：
2003
期刊：
影响因子：
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通讯作者：
O. Habuchi
中科院分区：
文献类型：
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作者：
T. Okuda;Toshihiko Sawada;H. Nakano;K. Matsubara;Y. Matsuda;M. Fukuta;O. Habuchi
N-乙酰半乳糖胺 4-磺基转移酶 (GalNAc4ST) 将硫酸盐转移至非还原性末端 GalNAc 残基的 4 位。我们之前克隆了人类 GalNAc4ST-1 cDNA。在本文中，我们报告了小鼠 GalNAc4ST-1 和 GalNAc4ST-2 的克隆、表征和染色体定位。小鼠 GalNAc4ST-1 和 GalNAc4ST-2 含有单个开放阅读框，可预测分别由 417 个和 413 个氨基酸残基组成的 II 型跨膜蛋白。两种异构体之间的氨基酸序列同一性为49%。当将cDNA转染至COS-7细胞时，对碳酸酐酶VI和GalNAcbeta1-4GlcNAcbeta1-辛基的磺基转移酶活性过表达，但对软骨素的磺基转移酶活性与对照水平相比没有增加。 Northern 印迹分析表明，GalNAc4ST-1 和 GalNAc4ST-2 的 2.4 kb 信息在肾脏中强烈表达，而这两种人类亚型在肾脏中几乎不表达。逆转录PCR分析表明，与人类GalNAc4ST-1不同，小鼠GalNAc4ST-1在垂体中的表达量仅为边缘水平，而GalNAc4ST-2在垂体中的表达量与肾脏中的表达量一样高。这些结果表明两种 GalNAc4ST 亚型的功能在人类和小鼠之间可能有所不同。通过荧光原位杂交，GalNAc4ST-1和GalNAc4ST-2基因分别定位于小鼠7B3染色体远端-B5近端和18A2染色体远端-B1近端。
N-Acetylgalactosamine 4-sulfotransferase (GalNAc4ST) transfers sulfate to position 4 of nonreducing terminal GalNAc residues. We previously cloned human GalNAc4ST-1 cDNA. In this paper, we report the cloning, characterization and chromosomal mapping of mouse GalNAc4ST-1 and GalNAc4ST-2. Mouse GalNAc4ST-1 and GalNAc4ST-2 contain single open reading frames that predict type II transmembrane proteins composed of 417 and 413 amino acid residues, respectively. The amino acid sequence identity between the two isoforms is 49%. When the cDNA was transfected to COS-7 cells, sulfotransferase activities toward carbonic anhydrase VI and GalNAcbeta1-4GlcNAcbeta1-octyl were overexpressed, but the sulfotransferase activity toward chondroitin showed no increase over the control level. Northern blot analysis showed that the 2.4 kb messages of GalNAc4ST-1 and GalNAc4ST-2 were strongly expressed in the kidney, where both of the human isoforms were hardly expressed. Reverse transcription-PCR analysis showed that, unlike human GalNAc4ST-1, the expression of mouse GalNAc4ST-1 in the pituitary gland was only marginal, while that of GalNAc4ST-2 in the pituitary gland was as high as that in the kidney. These results suggest that the functions of the two GalNAc4ST isoforms may differ between human and mouse. By fluorescence in situ hybridization, the GalNAc4ST-1 and GalNAc4ST-2 genes were localized to mouse chromosome 7B3 distal-B5 proximal and chromosome 18A2 distal-B1 proximal, respectively.