Development of a label-free assay for sodium-dependent phosphate transporter NaPi-IIb.
作者： Soo-Hang Wong , Alice Gao , Sabrina Ward , Charles Henley , Paul H. Lee
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摘要: The most widely used assay format for characterizing plasma membrane transporter activity measures accumulation of radiolabeled substrates in tissues or cells expressing the transporters. This had limitations and disadvantages; therefore, there was an unmet need development a homogeneous, nonradioactive proteins. In this report, authors describe label-free homogeneous sodium-dependent phosphate NaPi-IIb using Epic system. addition stimulated dynamic mass redistribution (DMR) profile unique to but not on parental cells. DMR specific because sulfate buffer alone did elicit same response. Furthermore, response observed sodium dependent, with Km values micromolar millimolar range, respectively. A known noncompetitive inhibitor shown completely inhibit phosphate-stimulated response, suggesting that is NaPi-IIb-mediated cellular event. results demonstrate novel developed studying transporter-mediated activity, platform could be applicable other
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sci-hub.se 参考文章(16)
Giustina Danisi, Heini Murer, Inorganic Phosphate Absorption in Small Intestine Comprehensive Physiology. pp. 323- 336 ,(2011) , 10.1002/CPHY.CP060412
M.L. Weiner, W.F. Salminen, P.R. Larson, R.A. Barter, J.L. Kranetz, G.S. Simon, Toxicological review of inorganic phosphates Food and Chemical Toxicology. ,vol. 39, pp. 759- 786 ,(2001) , 10.1016/S0278-6915(01)00028-X
Akira Matsuo, Tamotsu Negoro, Tomohisa Seo, Yuki Kitao, Masanori Shindo, Hiroko Segawa, Ken-ichi Miyamoto, Inhibitory effect of JTP-59557, a new triazole derivative, on intestinal phosphate transport in vitro and in vivo. European Journal of Pharmacology. ,vol. 517, pp. 111- 119 ,(2005) , 10.1016/J.EJPHAR.2005.05.003
Ji-Hu Zhang, Thomas D. Y. Chung, Kevin R. Oldenburg, A Simple Statistical Parameter for Use in Evaluation and Validation of High Throughput Screening Assays. Journal of Biomolecular Screening. ,vol. 4, pp. 67- 73 ,(1999) , 10.1177/108705719900400206
Eduardo Slatopolsky, Neal S. Bricker, The role of phosphorus restriction in the prevention of secondary hyperparathyroidism in chronic renal disease Kidney International. ,vol. 4, pp. 141- 145 ,(1973) , 10.1038/KI.1973.92
H. Hilfiker, O. Hattenhauer, M. Traebert, I. Forster, H. Murer, J. Biber, Characterization of a murine type II sodium-phosphate cotransporter expressed in mammalian small intestine Proceedings of the National Academy of Sciences of the United States of America. ,vol. 95, pp. 14564- 14569 ,(1998) , 10.1073/PNAS.95.24.14564
John A. Feild, Li Zhang, Kimberly A. Brun, David P. Brooks, Richard M. Edwards, Cloning and functional characterization of a sodium-dependent phosphate transporter expressed in human lung and small intestine. Biochemical and Biophysical Research Communications. ,vol. 258, pp. 578- 582 ,(1999) , 10.1006/BBRC.1999.0666
Hua Xu, Liqun Bai, James F. Collins, Fayez K. Ghishan, Molecular cloning, functional characterization, tissue distribution, and chromosomal localization of a human, small intestinal sodium-phosphate (Na+-Pi) transporter (SLC34A2). Genomics. ,vol. 62, pp. 281- 284 ,(1999) , 10.1006/GENO.1999.6009
Beatriz López-Corcuera, Rodrigo Martínez-Maza, Enrique Núñez, Michel Roux, Stéphane Supplisson, Carmen Aragón, Differential properties of two stably expressed brain-specific glycine transporters Journal of Neurochemistry. ,vol. 71, pp. 2211- 2219 ,(2002) , 10.1046/J.1471-4159.1998.71052211.X
Ye Fang, Label-free cell-based assays with optical biosensors in drug discovery. Assay and Drug Development Technologies. ,vol. 4, pp. 583- 595 ,(2006) , 10.1089/ADT.2006.4.583