The 5 angstrom projection structure of the transmembrane domain of the mannitol transporter enzyme II

The uptake of mannitol in Escherichia coli is controlled by the phosphoenol pyruvate dependent phosphotransferase system. Enzyme II mannitol (EIIMtl) i s part of the phosphotransferase system and consists of three covalently bo und domains. IICMtl, the integral membrane domain of EIIMtl, is responsible for mannitol transport across the cytoplasmic membrane. In order to unders tand this molecular process, two-dimensional crystals of IICMtl were grown by reconstitution into lipid bilayers and their structure was investigated by cryo-electron crystallography. The IICMtl crystals obey p22(1)2(1) symme try and have a unit cell of 125 Angstrom x 65 Angstrom, gamma = 90 degrees. A projection structure was determined at 5 Angstrom resolution using both electron images and electron diffractograms. The unit cell contains two IIC Mtl dimers with a size of about 40 Angstrom x 90 Angstrom which are oriente d up and down in the crystal. Each monomer exhibits six domains of high den sity which most likely correspond to transmembrane cl-helices and cytoplasm ic loops. (C) 1999 Academic Press.