Structural basis for detoxification and oxidative stress protection in membranes.

Synthesis of mediators of fever, pain and inflammation as well as protection against reactive molecules and oxidative stress is a hallmark of the MAPEG superfamily (membrane associated proteins in eicosanoid and glutathione metabolism). The structure of a MAPEG member, rat microsomal glutathione transferase 1, at 3.2 A resolution, solved here in complex with glutathione by electron crystallography, defines the active site location and a cytosolic domain involved in enzyme activation. The glutathione binding site is found to be different from that of the canonical soluble glutathione transferases. The architecture of the homotrimer supports a catalytic mechanism involving subunit interactions and reveals both cytosolic and membraneous substrate entry sites, providing a rationale for the membrane location of the enzyme.

[1]  Ralf Morgenstern,et al.  The 3-D structure of microsomal glutathione transferase 1 at 6 A resolution as determined by electron crystallography of p22(1)2(1) crystals. , 2002, Biochimica et biophysica acta.

[2]  Collaborative Computational,et al.  The CCP4 suite: programs for protein crystallography. , 1994, Acta crystallographica. Section D, Biological crystallography.

[3]  J. Thornton,et al.  PROCHECK: a program to check the stereochemical quality of protein structures , 1993 .

[4]  L. Ekström,et al.  Structural and Functional Aspects of Rat Microsomal Glutathione Transferase , 1997, The Journal of Biological Chemistry.

[5]  R. Morgenstern,et al.  Reactivity of cysteine-49 and its influence on the activation of microsomal glutathione transferase 1: evidence for subunit interaction. , 2000, Biochemistry.

[6]  Ralf Morgenstern,et al.  Bioinformatic and enzymatic characterization of the MAPEG superfamily , 2005, The FEBS journal.

[7]  G. Murshudov,et al.  Refinement of macromolecular structures by the maximum-likelihood method. , 1997, Acta crystallographica. Section D, Biological crystallography.

[8]  Kazushi Kimura,et al.  Implications of the aquaporin-4 structure on array formation and cell adhesion. , 2006, Journal of molecular biology.

[9]  Makoto Murakami,et al.  Regulation of Prostaglandin E2 Biosynthesis by Inducible Membrane-associated Prostaglandin E2 Synthase That Acts in Concert with Cyclooxygenase-2* , 2000, The Journal of Biological Chemistry.

[10]  Robert B Russell,et al.  A model for statistical significance of local similarities in structure. , 2003, Journal of molecular biology.

[11]  Randy Schekman,et al.  Protein Translocation Across Biological Membranes , 2005, Science.

[12]  R. Morgenstern,et al.  Microsomal glutathione transferase. Purification in unactivated form and further characterization of the activation process, substrate specificity and amino acid composition. , 1983, European journal of biochemistry.

[13]  B. Persson,et al.  Common structural features of mapeg—a widespread superfamily of membrane associated proteins with highly divergent functions in eicosanoid and glutathione metabolism , 2008, Protein science : a publication of the Protein Society.

[14]  R. MacKinnon,et al.  Principles of Selective Ion Transport in Channels and Pumps , 2005, Science.

[15]  A. Caccuri,et al.  Aggregation of pyrene-labeled microsomal glutathione S-transferase. Effect of concentration. , 1993, European journal of biochemistry.

[16]  H Hebert,et al.  The three‐dimensional map of microsomal glutathione transferase 1 at 6 Å resolution , 2000, The EMBO journal.

[17]  J. Zou,et al.  Improved methods for building protein models in electron density maps and the location of errors in these models. , 1991, Acta crystallographica. Section A, Foundations of crystallography.

[18]  R. Armstrong,et al.  Spectroscopic and kinetic evidence for the thiolate anion of glutathione at the active site of glutathione S-transferase. , 1989, Biochemistry.

[19]  H. Jörnvall,et al.  Activation of rat liver microsomal glutathione transferase by limited proteolysis. , 1989, The Biochemical journal.

[20]  R. Armstrong,et al.  Structure, catalytic mechanism, and evolution of the glutathione transferases. , 1997, Chemical research in toxicology.

[21]  Ralf Morgenstern,et al.  Human Microsomal Prostaglandin E Synthase-1 , 2003, Journal of Biological Chemistry.

[22]  R. Morgenstern,et al.  Parameters for the two-dimensional crystallization of the membrane protein microsomal glutathione transferase. , 1998, Journal of structural biology.

[23]  R. Armstrong,et al.  Stress sensor triggers conformational response of the integral membrane protein microsomal glutathione transferase 1. , 2004, Biochemistry.

[24]  G L Gilliland,et al.  The three-dimensional structure of a glutathione S-transferase from the mu gene class. Structural analysis of the binary complex of isoenzyme 3-3 and glutathione at 2.2-A resolution. , 1992, Biochemistry.

[25]  S. Harrison,et al.  Lipid–protein interactions in double-layered two-dimensional AQP0 crystals , 2005, Nature.

[26]  B. Wallace,et al.  HOLE: a program for the analysis of the pore dimensions of ion channel structural models. , 1996, Journal of molecular graphics.

[27]  S. Colowick,et al.  Methods in Enzymology , Vol , 1966 .

[28]  W. Pearson.  Phylogenies of glutathione transferase families. , 2005, Methods in enzymology.

[29]  R. Morgenstern.  A simple alternate substrate test can help determine the aqueous or bilayer location of binding sites for hydrophobic ligands/substrates on membrane proteins. , 1998, Chemical Research in Toxicology.

[30]  Andreas Engel,et al.  Structural determinants of water permeation through aquaporin-1 , 2000, Nature.

[31]  P E Bourne,et al.  Protein structure alignment by incremental combinatorial extension (CE) of the optimal path. , 1998, Protein engineering.

[32]  D. Maskell,et al.  Toward a structural understanding of the dehydratase mechanism. , 2002, Structure.

[33]  R. Armstrong,et al.  Kinetic analysis of the slow ionization of glutathione by microsomal glutathione transferase MGST1. , 2001, Biochemistry.

[34]  Y. Fujiyoshi,et al.  Trehalose embedding technique for high-resolution electron crystallography: application to structural study on bacteriorhodopsin. , 1999, Journal of electron microscopy.

[35]  B. Mannervik,et al.  Synthesis and characterization of 6-chloroacetyl-2-dimethylaminonaphthalene as a fluorogenic substrate and a mechanistic probe for glutathione transferases. , 2002, Analytical Biochemistry.

[36]  A. Oakley.  Glutathione transferases: new functions. , 2005, Current opinion in structural biology.

[37]  K. Austen,et al.  Site-directed Mutagenesis of Human Leukotriene C4Synthase* , 1997, The Journal of Biological Chemistry.

[38]  R A Crowther,et al.  MRC image processing programs. , 1996, Journal of structural biology.

[39]  Yoshinori Fujiyoshi,et al.  Development of a superfluid helium stage for high-resolution electron microscopy , 1991 .

[40]  Yoshinori Fujiyoshi,et al.  Atomic model of plant light-harvesting complex by electron crystallography , 1994, Nature.

[41]  Kazutoshi Tani,et al.  Improved specimen preparation for cryo-electron microscopy using a symmetric carbon sandwich technique. , 2004, Journal of structural biology.

[42]  R. Morgenstern,et al.  Microsomal glutathione S-transferase. Purification, initial characterization and demonstration that it is not identical to the cytosolic glutathione S-transferases A, B and C. , 2005, European journal of biochemistry.

[43]  C. Sander,et al.  Protein structure comparison by alignment of distance matrices. , 1993, Journal of molecular biology.

[44]  K. Ruan,et al.  Topology of prostaglandin H synthase-1 in the endoplasmic reticulum membrane. , 1995, Archives of biochemistry and biophysics.

[45]  G J Mulder,et al.  Studies on the activity and activation of rat liver microsomal glutathione transferase with a series of glutathione analogues. , 1991, The Journal of biological chemistry.

[46]  W. Kühlbrandt,et al.  Human leukotriene C(4) synthase at 4.5 A resolution in projection. , 2004, Structure.

[47]  R. Morgenstern,et al.  Evidence that rat liver microsomal glutathione transferase is responsible for glutathione-dependent protection against lipid peroxidation. , 1993, Biochemical pharmacology.