Marine pollution is of concern for human health through our exposure to contaminated food from the sea. What remains poorly understood is why some chemicals are persistent, accumulating in marine organisms and then in humans, while others are not. Multidrug resistance (MDR) transporters, belonging to the ATP Binding Cassette (ABC) family, are major biological determinants of intracellular chemical accumulation. While they have been implicated as determinants of environmental chemical persistence and used as tools for predicting availability and efficacy of drugs, they have yet to be systematically applied to predicting persistence of pollutants. The investigators'preliminary data indicate striking functional conservation of the major sub-family types (ABCB, ABCC and ABCG) of xenobiotic eliminating transporters between sea urchins and man. This application explores the molecular basis for this conserved substrate selectivity as a first step towards application of transporter biology to prediction of pollutant persistence. In this project, the investigators will over-express, solubilize, and purify sea urchin multidrug efflux transporter proteins, develop assays for their interaction with major marine pollutants and attempt to determine their high-resolution structures. They will measure their interaction with persistent marine pollutants using anisotropy, ATPase and whole cell assays. By comparing structure and functions of sea urchin with those of mammalian transporter proteins, already available through the TransportPDB pipeline, they will identify conserved residues and structural features that are essential for predicting substrate interaction in the poly-specific binding pocket of these ABC transporters

Public Health Relevance

Certain molecular transporters found in sea urchins and other marine organisms determine the uptake of persistent pollutants from the sea. The goal of this high-impact project is to provide a detailed molecular framework for understanding the structure and function of these important transporters. This application fulfills an important need to develop biologically-based tools to predict bioaccumulation and toxicity of marine pollutants as relevant to human health and environment.

Agency
National Institute of Health (NIH)
Type
Research Project (R01)
Project #
5R01ES021985-03
Application #
8707456
Study Section
Special Emphasis Panel (ZES1)
Program Officer
Tyson, Frederick L
Project Start
Project End
Budget Start
Budget End
Support Year
3
Fiscal Year
2014
Total Cost
Indirect Cost
Name
University of California San Diego
Department
Zoology
Type
Earth Sciences/Resources
DUNS #
City
La Jolla
State
CA
Country
United States
Zip Code
92093
Gökirmak, Tufan; Shipp, Lauren E; Campanale, Joseph P et al. (2014) Transport in technicolor: mapping ATP-binding cassette transporters in sea urchin embryos. Mol Reprod Dev 81:778-93
Campanale, Joseph P; Gökirmak, Tufan; Espinoza, Jose A et al. (2014) Migration of sea urchin primordial germ cells. Dev Dyn 243:917-27
Ward, Andrew B; Szewczyk, Paul; Grimard, Vinciane et al. (2013) Structures of P-glycoprotein reveal its conformational flexibility and an epitope on the nucleotide-binding domain. Proc Natl Acad Sci U S A 110:13386-91