Structure-based analysis of CysZ-mediated cellular uptake of sulfate
Zahra Assur Sanghai,
Qun Liu,
Oliver B Clarke,
Meagan Belcher-Dufrisne,
Pattama Wiriyasermkul,
M Hunter Giese,
Edgar Leal-Pinto,
Brian Kloss,
Shantelle Tabuso,
James Love,
Marco Punta,
Surajit Banerjee,
Kanagalaghatta R Rajashankar,
Burkhard Rost,
Diomedes Logothetis,
Matthias Quick,
Wayne A Hendrickson,
Filippo Mancia
Affiliations
Zahra Assur Sanghai
Department of Physiology and Cellular Biophysics, Columbia University, New York, United States; Department of Biochemistry and Molecular Biophysics, Columbia University, New York, United States
Qun Liu
Biology Department, Brookhaven National Laboratory, Upton, United States
Department of Biochemistry and Molecular Biophysics, Columbia University, New York, United States
Meagan Belcher-Dufrisne
Department of Physiology and Cellular Biophysics, Columbia University, New York, United States
Pattama Wiriyasermkul
Center for Molecular Recognition, Department of Psychiatry, Columbia University, New York, United States
M Hunter Giese
Department of Physiology and Cellular Biophysics, Columbia University, New York, United States; Department of Biochemistry and Molecular Biophysics, Columbia University, New York, United States
Edgar Leal-Pinto
Department of Physiology and Biophysics, Virginia Commonwealth University School of Medicine, Richmond, United States; Department of Pharmaceutical Sciences, School of Pharmacy, Bouvé College of Health Sciences, Northeastern University, Boston, United States
Brian Kloss
New York Structural Biology Center, New York, United States
Shantelle Tabuso
New York Structural Biology Center, New York, United States
James Love
New York Structural Biology Center, New York, United States
Marco Punta
Centre for Evolution and Cancer, The Institute of Cancer Research, London, United Kingdom
Surajit Banerjee
Department of Chemistry and Chemical Biology, Cornell University, NE-CAT, Argonne, United States
Kanagalaghatta R Rajashankar
Department of Chemistry and Chemical Biology, Cornell University, NE-CAT, Argonne, United States
Burkhard Rost
Department of Informatics, Technical University of Munich, Munich, Germany
Diomedes Logothetis
Department of Physiology and Biophysics, Virginia Commonwealth University School of Medicine, Richmond, United States; Department of Pharmaceutical Sciences, School of Pharmacy, Bouvé College of Health Sciences, Northeastern University, Boston, United States
Matthias Quick
Center for Molecular Recognition, Department of Psychiatry, Columbia University, New York, United States; Division of Molecular Therapeutics, New York State Psychiatric Institute, New York, United States
Wayne A Hendrickson
Department of Physiology and Cellular Biophysics, Columbia University, New York, United States; Department of Biochemistry and Molecular Biophysics, Columbia University, New York, United States; New York Structural Biology Center, New York, United States
Sulfur, most abundantly found in the environment as sulfate (SO42-), is an essential element in metabolites required by all living cells, including amino acids, co-factors and vitamins. However, current understanding of the cellular delivery of SO42- at the molecular level is limited. CysZ has been described as a SO42- permease, but its sequence family is without known structural precedent. Based on crystallographic structure information, SO42- binding and flux experiments, we provide insight into the molecular mechanism of CysZ-mediated translocation of SO42- across membranes. CysZ structures from three different bacterial species display a hitherto unknown fold and have subunits organized with inverted transmembrane topology. CysZ from Pseudomonas denitrificans assembles as a trimer of antiparallel dimers and the CysZ structures from two other species recapitulate dimers from this assembly. Mutational studies highlight the functional relevance of conserved CysZ residues.
