Positioning membrane proteins by novel protein engineering and biophysical approaches
Abbreviated Journal Title
J. Mol. Biol.
membrane protein; orientation; insertion; polarized FTIR; fluorescence; quenching; RELAXATION MAGNETIC-RESONANCE; PANCREATIC PHOSPHOLIPASE A(2); REFLECTION; FTIR SPECTROSCOPY; LIPID BILAYERS; X-RAY; INFRARED-SPECTROSCOPY; NMR-SPECTROSCOPY; GATING MECHANISM; ORIENTATION; BINDING; Biochemistry & Molecular Biology
Membrane proteins are unique, in that they can function properly only when they are bound to cellular membranes in a distinct manner. Therefore, positioning of, membrane proteins with respect to the membrane is required in addition to the three-dimensional structures in order to understand their detailed molecular mechanisms. Atomic-resolution structures of membrane proteins that have been determined to date provide the atom coordinates in arbitrary coordinate systems with no relation to the membrane and therefore provide little or no information on how the protein would interact with the membrane. This is especially true for peripheral membrane proteins, because they, unlike integral proteins, are devoid of well-defined hydrophobic transmembrane domains. Here, we present a novel technique for determination of the configuration of a protein-membrane complex that involves protein ligation, segmental isotope labeling, polarized infrared spectroscopy, membrane depth dependent fluorescence quenching, and analytical geometry algorithms. We have applied this approach to determine the structure of a membranebound phospholipase A(2). Our results provide an unprecedented structure of a membrane-bound protein in which the z-coordinate of each atom is the distance from the membrane center and therefore allows precise location of each amino acid relative to the membrane. Given the functional significance of the orientation and location of membrane-bound proteins with respect to the membrane, we propose to specify this structural feature as the "quinary" structure of membrane proteins. (c) 2005 Elsevier Ltd. All rights reserved.
Journal of Molecular Biology
"Positioning membrane proteins by novel protein engineering and biophysical approaches" (2005). Faculty Bibliography 2000s. 5716.