Abstract
May 3, 2006
Cornell University Biophysics Colloquium
700 Clark Hall, 4:30 pm


“A two billion year old tale of membrane transport. It's a gas!”

Robert M. Stroud
Professor of Biochemistry & Biophysics
Professor of Pharmaceutical Chemistry
Department of Biochemistry & Biophysics
University of California San Francisco
Website

The families of proteins that transport water, glycerol, ammonia and carbon dioxide are found across all species. Ammonia transporters and Aquaporins are highly selective transmembrane channels for permeation of specific small molecules, with absolute exclusion of ions including protons or OH- anions and charged solutes, and without dissipating the electrochemical potential across the cell membrane. The mechanisms of transport are deduced from structures and assays. The E.coli glycerol facilitator (GlpF), a water and a glycerol conducting channel structure is determined at 1.9Å resolution1,2. Subsequently a structure for AQP1 from erythrocytes followed, and we determined structures for AQP04, AQPZ3, and AQPM. These channels will be compared to illustrate their function. A line of hydrogen bonded water molecules are hydrogen bonded to a line of 8 hydrogen bond acceptors. The first structure of an ammonia transporter from the Amt/MEP/Rh superfamily, determined to 1.35Å resolution5 shows it to be a channel that spans the membrane 11 times. Two structurally similar halves span the membrane with opposite polarity. Structures with and without ammonia, or methyl ammonia show a vestibule that recruits NH4+/NH3, a binding site for NH4+ and a 20Å long hydrophobic channel that lowers the NH4+ pKa to below 6 and conducts NH3. Favorable interactions for NH3 are seen within the channel and use conserved histidines. Reconstitution of AmtB into vesicles shows that AmtB conducts uncharged NH3. These are the first structures of gas channels!

1. Fu, D., Libson, A., Miercke, L., Weitzman, C., Nollert, P., Krucinski, J., and Stroud, R. (2000). Science 290: 481-486. The Structure of a Glycerol Conducting Channel Reveals the Basis for its Selectivity.
2. Tajkhorshid, E. Nollert, P. Jensen, M.O. Miercke, L.J.W. O’Connell, J. Stroud, R.M. Schulten, K. (2002) Science 296, 525-530. Control of the Selectivity of the Aquaporin Water Channel Family by Global Orientational Tuning
3. Savage, D. F., Egea, P.F., Robles, Y.C., O’Connell III, J.D., and Stroud, R.M. (2003) PLoS Biology 1 334-340 with cover, and Synopsis 1 302. Architecture and selectivity in aquaporins: 2.5Å X-ray structure of aquaporin Z.
4. Harries, W. E., Akhavan, D., Miercke, L. J., Khademi, S., Stroud, R. M. The channel architecture of aquaporin 0 at a 2.2-Å resolution. Proc Natl Acad Sci U S A. 101, 14045-14050
5. Khademi, S., O'Connell, J., 3rd, Remis, J., Robles-Colmenares, Y., Miercke, L. J., Stroud, R. M. Mechanism of ammonia transport by Amt/MEP/Rh: structure of AmtB at 1.35 A Science 305 1587-94

Return to Biophysics Colloquia page.