Molecular cloning and characterization of NEU4, the fourth member of the human sialidase gene family

被引:97
|
作者
Monti, E
Bassi, MT
Bresciani, R
Civini, S
Croci, GL
Papini, N
Riboni, M
Zanchetti, G
Ballabio, A
Preti, A
Tettamanti, G
Venerando, B
Borsani, G
机构
[1] Univ Brescia, Dept Biomed Sci & Biotechnol, I-25123 Brescia, Italy
[2] IRCCS E Medea, I-23842 Bosisio Parini, Lecco, Italy
[3] LITA, Dept Med Chem Biochem & Biotechnol, I-20090 Milan, Italy
[4] FIRC Inst Mol Oncol, I-20139 Milan, Italy
[5] Telethon Inst Genet & Med, I-80131 Naples, Italy
[6] Univ Naples 2, Fac Med, Naples, Italy
关键词
human sialidase; gene structure; gene expression profile; transient transfection; subcellular localization;
D O I
10.1016/j.ygeno.2003.08.019
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Several mammalian sialidases have been cloned so far and here we describe the identification and expression of a new member of the human sialidase gene family. The NEU4 gene, identified by searching sequence databases for entries showing homologies to the human cytosolic sialidase NEU2, maps in 2q37 and encodes a 484-residue protein. The polypeptide contains all the typical sialidase amino acid motifs and, apart from an amino acid stretch that appears unique among mammalian sialidases, shows a high degree of homology for NEU2 and the plasma membrane-associated (NEU3) sialidases. RNA dot-blot analysis showed a low but wide expression pattern, with the highest level in liver. Transient transfection in COS7 cells allowed the detection of a sialidase activity toward the artificial substrate 4MU-NeuAc in the acidic range of pH. Immunofluorescence staining and Western blot analysis demonstrated the association of NEU4 with the inner cell membranes. (C) 2003 Elsevier Inc. All rights reserved.
引用
收藏
页码:445 / 453
页数:9
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