The aim of this proposal is to determine the structure of AdiC, a member of a widespread and little-understood class of membrane transport proteins - the APC superfamily. APC proteins, nearly ubiquitous in the biological world, are used to move amino acids, polyamines, and a variey of organic cations across cell membranes for a multitude of physiological purposes. Although this is a huge molecular superfamily, no high-resolution structures are known for any APC proteins, and our understanding of their fundamental mechanisms of substrate transport is currently shallow. The bacterial APC protein AdiC, which moves arginine into many enteric bacteria, is a key player in the """"""""extreme acid resistance"""""""" response that these organisms use to survive exposure to the acid environment of the stomach. This particular APC protein may be overexpressed, purified, functionally reconstituted in artificial membranes, and, most importantly, crystallized. The project aims to determine the structure of AdiC by x-ray crystallography, and to use this structural information to understand how this protein specifically recognizes its organocationic substrates and how it moves threse across the biological membrane.
This project is relevant to human health in two ways. First, by providing fundamental information on the edge of discovery about the molecular workings of a new class of membrane protein, it will illuminate the ways that cells nourish themselves with essential amino acid nutrients. Second, this particular protein family is intimately involved not only in essential human physiologies in health, but also in devastating pathologies, in particular a variety of cancers that require APC transporters to supply amino acids to fast-growing, malignant cells.
|Tsai, Ming-Feng; Miller, Christopher (2013) Substrate selectivity in arginine-dependent acid resistance in enteric bacteria. Proc Natl Acad Sci U S A 110:5893-7|
|Tsai, Ming-Feng; McCarthy, Patrick; Miller, Christopher (2013) Substrate selectivity in glutamate-dependent acid resistance in enteric bacteria. Proc Natl Acad Sci U S A 110:5898-902|
|Tsai, Ming-Feng; Fang, Yiling; Miller, Christopher (2012) Sided functions of an arginine-agmatine antiporter oriented in liposomes. Biochemistry 51:1577-85|
|Lim, Hyun-Ho; Fang, Yiling; Williams, Carole (2011) High-efficiency screening of monoclonal antibodies for membrane protein crystallography. PLoS One 6:e24653|
|Miller, Christopher (2010) CFTR: break a pump, make a channel. Proc Natl Acad Sci U S A 107:959-60|
|Theobald, Douglas L; Miller, Christopher (2010) Membrane transport proteins: surprises in structural sameness. Nat Struct Mol Biol 17:2-3|
|Fang, Yiling; Jayaram, Hariharan; Shane, Tania et al. (2009) Structure of a prokaryotic virtual proton pump at 3.2 A resolution. Nature 460:1040-3|