Name: Prachee Bele
Diploma / M.Sc degree: University of Hyderabad, India
(June 2009)

PhD Project: Cloning,expression and Characterization of hMRP3,hABCG2 and pAbcg2 in Pichia pastoris and a detailed study of each, in artificial Liposomes


Homepage: http://www.uni-muenster.de/Chemie.bc/staff/bc-staff.html

Abstract of Research Project

ABC transporters constitute one of the largest and ancient families of the membrane proteins. Especially their role in the Multidrug Resistance has gained considerable attention of scientists across the world. So far 48 ABC transporters have been discovered in human. While some of them have already been studied, some are yet a mystery and need further study. We are working on the functional aspects of two such proteins namely the human ABCC3 and the porcine Abcg2 involved in the Multidrug resistance at the blood brain barrier or in cancer cells. Although both the proteins involved in the similar process, efflux of the drug out of the cell, their structures vary altogether. While Abcg2 is a half transporter with only a single TMD (Trans Membrane Domain) and NBD (Nucleotide Binding Domain), ABCC3 is a full transporter having 2 TMDs , 2NBDs along with an additional TMD domain.

In our lab, we are expressing both the proteins and different domains of hABCC3 separately, by using yeast Pichia pastoris as the host system for expression. The expressed, functional protein/domain will then be studied in artificial liposomes of different lipid constitution, for their activity with different anticancer drugs as substrates. The activity measurements will be performed by various methods such as ATPase assay, radiolabelled substrates or scintillation assays. In order to study the structure-function relationship, cooperativity and substrate specificity of these proteins and their domains in detail, we will reconstitute the full transporter, the different domains and combinations of domains also obtained from other transporters. The half transporter pAbcg2 is planned to be combined with TMDS of other full transporters.



Publications

J. Lemmen, I. E. P. Tozakidis, P. Bele, H.-J. Galla
Constitutive androstane receptor upregulates Abcb1 and Abcg2 at the blood–brain barrier after CITCO activation
Brain Research 1501 (2013), 68-80.


Prachee Bele
eMail: Prachee Bele