C-terminal splicing of NTPDase2 provides distinctive catalytic properties, cellular distribution and enzyme regulation.
C-terminal splicing of NTPDase2 provides distinctive catalytic properties, cellular distribution and enzyme regulation.
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NTPDase2 的 C 端剪接提供了独特的催化特性、细胞分布和酶调节。
DOI：
10.1042/bj20040852
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发表时间：
2005
期刊：
影响因子：
--
通讯作者：
P. Thorne
中科院分区：
文献类型：
--
作者：
Carol J. H. Wang;S. Vlajkovic;G. Housley;N. Braun;H. Zimmermann;S. Robson;J. Sévigny;C. Soeller;P. Thorne
本研究提供了选择性剪接的NTPDase 2（胞外核苷三磷酸二磷酸水解酶-2）参与调节细胞外核苷酸浓度在一系列器官系统的功能表征。通过大鼠NTPDase 2基因的选择性剪接产生的新型NTPDase 2 β同种型提供了延长的细胞内C-末端，并在获得几个细胞内蛋白激酶CK 2（酪蛋白激酶2）磷酸化位点和失去细胞内蛋白激酶C基序方面与NTPDase 2 α同种型区分开来。用含有NTPDase 2 α或NTPDase 2 β cDNA的质粒稳定转染中国仓鼠卵巢S细胞。成像研究表明，NTPDase 2 α主要是膜结合，而NTPDase 2 β结合细胞表面和细胞内定位。α和β亚型对核苷-5 '-三磷酸和核苷-5'-二磷酸的二价阳离子依赖性和底物特异性存在差异。NTPD酶2 β表现出ATP酶活性降低，无明显的ATP酶活性。NTPD酶2亚型对抑制剂如苏拉明和磷酸吡哆醛-6-偶氮苯基-2 '，4'-二磺酸表现出相似的敏感性，并通过蛋白激酶进行差异调节。NTPDase 2 β被细胞内蛋白激酶CK 2磷酸化上调，而NTPDase 2 α活性被蛋白激酶C磷酸化下调。结果表明，交替编码的细胞内C-末端结构域贡献独特的表型变异，相对于细胞外核苷酸特异性，水解动力学，蛋白激酶依赖的细胞内调节和蛋白质运输。这些发现通过表征C-末端结构域对许多酶的特征性质的贡献来推进该酶系统的分子生理学。
The present study provides functional characterization of alternative splicing of the NTPDase2 (ecto-nucleoside triphosphate diphosphohydrolase-2) involved in the regulation of extracellular nucleotide concentrations in a range of organ systems. A novel NTPDase2beta isoform produced by alternative splicing of the rat NTPDase2 gene provides an extended intracellular C-terminus and distinguishes itself from NTPDase2alpha isoform in gaining several intracellular protein kinase CK2 (casein kinase 2) phosphorylation sites and losing the intracellular protein kinase C motif. The plasmids containing NTPDase2alpha or NTPDase2beta cDNA were used to stably transfect Chinese-hamster ovary-S cells. Imaging studies showed that NTPDase2alpha was predominantly membrane-bound, whereas NTPDase2beta had combined cell surface and intracellular localization. alpha and beta isoforms showed variations in divalent cation dependence and substrate specificity for nucleoside-5'-triphosphates and nucleoside-5'-diphosphates. NTPDase2beta exhibited reduced ATPase activity and no apparent ADPase activity. NTPDase2 isoforms demonstrated similar sensitivity to inhibitors such as suramin and pyridoxal phosphate-6-azophenyl-2',4'-disulphonic acid, and differential regulation by protein kinases. NTPDase2beta was up-regulated by intracellular protein kinase CK2 phosphorylation, whereas NTPDase2alpha activity was down-regulated by protein kinase C phosphorylation. The results demonstrate that alternative coding of the intracellular C-terminal domain contributes distinctive phenotypic variation with respect to extracellular nucleotide specificity, hydrolysis kinetics, protein kinase-dependent intracellular regulation and protein trafficking. These findings advance the molecular physiology of this enzyme system by characterizing the contribution of the C-terminal domain to many of the enzyme's signature properties.
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影响因子：
13.8
作者：
Abbracchio, MP;Boeynaems, JM;Burnstock, G
通讯作者：
Burnstock, G
影响因子：
21.1
作者：
B. Khakh;G. Burnstock;C. Kennedy;B. King;R. North;P. Séguéla;M. Voigt;P. Humphrey
通讯作者：
B. Khakh;G. Burnstock;C. Kennedy;B. King;R. North;P. Séguéla;M. Voigt;P. Humphrey
DOI：
10.1046/j.1432-1033.2001.01896.x
发表时间：
2001
期刊：
European journal of biochemistry
影响因子：
--
作者：
P. Heine;N. Braun;Jean Sévigny;Simon C. Robson;Jörg Servos;Herbert Zimmermann
通讯作者：
P. Heine;N. Braun;Jean Sévigny;Simon C. Robson;Jörg Servos;Herbert Zimmermann
鉴定垂体和耳蜗中通过选择性剪接产生的 P2xR1-嘌呤受体亚基的短形式。
影响因子：
3.1
作者：
Housley,GD;Greenwood,D;Bennett,T;Ryan,AF
通讯作者：
Ryan,AF
通过寡聚状态控制细胞膜外源 ATP 酶：分子间交联调节 ATP 酶活性。
影响因子：
2.9
作者：
Stout,JG;Kirley,TL
通讯作者：
Kirley,TL