Premium
Assembly and Application of Phospholipid Nanodiscs for the Study of P‐Glycoprotein
Author(s) -
McClean Mindy,
Brewer Frances,
Wise John,
Vogel Pia
Publication year - 2015
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.29.1_supplement.721.24
Subject(s) - nanodisc , p glycoprotein , chemistry , mutant , membrane protein , computational biology , multiple drug resistance , biochemistry , biology , gene , membrane , antibiotics
As many as 40% of recurring cancers often develop multidrug resistance (MDR) to chemotherapeutic treatment due to over‐expression of P‐glycoprotein (P‐gp), a member of the ABC transporter protein family. Our research has focused on finding inhibitors of P‐gp that are capable of re‐sensitizing MDR cancers to therapy. In order to investigate the mechanism of action of the discovered inhibitors, we employ electron spin resonance spectroscopy and biochemical assays. Due to the inherent instability of these mammalian membrane proteins, many of these analyses are challenging. We therefore are establishing nanodisc technology to incorporate and stabilize purified P‐gp for our drug finding and mechanistic studies. Nanodiscs are phospholipid bilayers contained within a membrane scaffold protein and mimic native membrane conditions. Initial studies indicated that a cysteine‐less mutant of the mouse MDR3 P‐gp could be incorporated into nanodiscs and showed enhanced catalytic activity and stability. We are currently establishing nanodiscs as a uniform platform from which different mutants of P‐pg can be closely studied and characterized. Nanodiscs may also enable us to utilize human MDR1 P‐gp in drug finding studies. This particular protein has been shown to be too unstable in micelle solution to be successfully used in these experiments. This work is supported by NIH NIGMS [Grant 1R15‐GM094771‐01A1] to PDV and JGW, SMU Research Council, Dean's Research Council, and CD4, and the Communities Foundation of Texas, Dallas. Undergraduate support was received from the Hamilton Family, Dallas, SMU Undergraduate Research Assistantship, and Engaged Learning Fellowship.