geoscience.net logo
+ Resolve Article
+ Follow Us
Follow on FacebookFollow on Facebook
Follow on TwitterFollow on Twitter

+ Translate
+ Subscribe to Site Feed
GeoScience Most Shared ContentMost Shared Content

Drug inhibition of Gly-Sar uptake and hPepT1 localization using hPepT1-GFP fusion protein


, : Drug inhibition of Gly-Sar uptake and hPepT1 localization using hPepT1-GFP fusion protein. Aaps Pharmsci 3(1): E2-E2

An hPepT1-GFP fusion construct was made to study drug inhibition of dipeptide uptake and apical, basolateral, or subcellular hPepT1 localization. The hPepT1 stop codon was mutated by polymerase chain reaction and was subsequently cloned into the pEGFP-N1 vector. The hPepT1-GFP fusion construct was then transfected into Caco-2 and HeLa cells, and drug inhibition was studied by inhibiting 3H-Gly-Sar uptake. Western blot analysis was used to determine hPepT1-GFP expression levels and confocal microscopy was used to examine the localization. Both anti-hPepT1 antibody and anti-GFP antibody recognized a 120-kd hPepT1-GFP fusion protein in the transfected cells. The 3H-Gly-Sar uptake in transfected HeLa cells was enhanced more than 20 times compared with the control. Valacyclovir (5 mmol/L) was able to completely inhibit 3H-Gly-Sar uptake in these transfected cells. Confocal microscopy showed that the hPepT1-GFP mainly localized in the Caco-2 cell apical membrane, but was present throughout the entire HeLa cell membranes. The hPepT1-GFP fusion protein was not found in either early endosome or lysosome of Caco-2 cells under normal conditions; however, it was detected in some subsets of lysosomes and early endosomes in phorbol 12-myristate 13-acetate (PMA)-treated Caco-2 cells.

US$29.90

PMID: 11741253


Other references

Rios, A.; Kavuluru, R., 2013: Supervised Extraction of Diagnosis Codes from EMRs: Role of Feature Selection, Data Selection, and Probabilistic Thresholding. Extracting diagnosis codes from medical records is a complex task carried out by trained coders by reading all the documents associated with a patient's visit. With the popularity of electronic medical records (EMRs), computational approaches...

Godoy A.C.; Garza S.H.; Huitron R.M.V., 1998: Growth and sugar accumulation in grape (Vitis vinifera L.) fruits under different water conditions. Trials were carried out in 1994 in an 18-year-old vineyard in Coahuila, Mexico, planted with the Grenache cultivar, which was subjected during phase III of fruit growth to 3 evapotranspiration (ET) irrigation levels (0, 30 and 70%). Fruit volume a...

de Haydu, C.; Black, J.D.; Schwab, C.L.; English, D.P.; Santin, A.D., 2016: An update on the current pharmacotherapy for endometrial cancer. Endometrial cancer (EC) is the most common gynecologic malignancy in the developed world and is increasing in incidence. While the mainstay of treatment for EC is surgery followed by chemotherapy and/or radiation therapy, the available pharmacothe...

Jaffe, K.; Ramos, C.; Issa, S., 1995: Trophic interactions between ants and termites that share common nests. The neotropical termite Nasutitermes corniger (Motschulsky) cohabits with various ants species. During the wet season, ants use part of the termite nest as shelter and may prey on live or dead, or both, termite workers, while providing the termite...

Erben, H.K., 1969: Faunenprovinzielle Beziehungen zwischen unterdevonischen Trilobiten der Karnischen Alpen and Zentralasiens. Carinthia II (Sonderheft): 27: 19-20

Nagral, S., 2015: Mlk. Indian Journal of Medical Ethics 12(1): 61-62

Dalvi, J.M.; Chen, L., 2003: Ionic conjugated polymer Synthesis and biosensor application. Abstracts of Papers American Chemical Society 225(1-2): PMSE 184

Inga M.Jacobs, 2012: A community in the Orange the development of a multi-level water governance framework in the Orange-Senqu River basin in Southern Africa. An ensemble of normative codes of conduct in the form of global, regional and domestic norms, principles of best practice and laws have developed over time providing standards of appropriate behaviour in the governance of transboundary rivers in a...

Chernin, M.M.; Amberg, J.R.; Kogan, F.J.; Morgan, T.R.; Sampliner, R.E., 1986: Efficacy of radiologic studies in the detection of Barrett's esophagus. Esophageal radiography using two different air-contrast techniques was used to examine 30 patients with Barrett's esophagus and 18 controls. All patients had upper gastrointestinal endoscopy and biopsy of the esophagus. The radiographs were r...

Khundadze G.R.; Tsanava G.M.; Goroziana C.V.; Mamamtavrishvili O.G.; Mikeladze T.D., 1970: The use of viadril g in anesthesiological practice. Trudy Tbilisskogo Instituta Usovershenstvovaniya Vrachei 11: 56-58