The C-terminal fragment of the ribosomal P protein complexed to trichosanthin reveals the interaction between the ribosome-inactivating protein and the ribosome

Ribosome-inactivating proteins (RIPs) inhibit protein synthesis by enzymatically depurinating a specific adenine residue at the sarcin-ricin loop of the 28S rRNA, which thereby prevents the binding of elongation factors to the GTPase activation centre of the ribosome. Here, we present the 2.2 Å crystal structure of trichosanthin (TCS) complexed to the peptide SDDDMGFGLFD, which corresponds to the conserved C-terminal elongation factor binding domain of the ribosomal P protein. The N-terminal region of this peptide interacts with Lys173, Arg174 and Lys177 in TCS, while the C-terminal region is inserted into a hydrophobic pocket. The interaction with the P protein contributes to the ribosome-inactivating activity of TCS. This 11-mer C-terminal P peptide can be docked with selected important plant and bacterial RIPs, indicating that a similar interaction may also occur with other RIPs.

[1]  P. Shaw,et al.  The C-terminal end of P proteins mediates ribosome inactivation by trichosanthin but does not affect the pokeweed antiviral protein activity. , 2008, Biochemical and biophysical research communications.

[2]  J. Gariépy,et al.  The catalytic subunit of shiga-like toxin 1 interacts with ribosomal stalk proteins and is inhibited by their conserved C-terminal domain. , 2008, Journal of molecular biology.

[3]  Narayanan Eswar,et al.  Structure of the mammalian 80S ribosome at 8.7 A resolution. , 2008, Structure.

[4]  J. Piccirilli,et al.  Evidence for the importance of electrostatics in the function of two distinct families of ribosome inactivating toxins. , 2007, RNA.

[5]  G. Zhu,et al.  Interaction between trichosanthin, a ribosome-inactivating protein, and the ribosomal stalk protein P2 by chemical shift perturbation and mutagenesis analyses , 2007, Nucleic acids research.

[6]  M. Akke,et al.  The ribosomal stalk binds to translation factors IF2, EF-Tu, EF-G and RF3 via a conserved region of the L12 C-terminal domain. , 2007, Journal of molecular biology.

[7]  J. Piccirilli,et al.  The electrostatic character of the ribosomal surface enables extraordinarily rapid target location by ribotoxins , 2006, Nature Structural &Molecular Biology.

[8]  J. Frank,et al.  Interaction of the G' domain of elongation factor G and the C-terminal domain of ribosomal protein L7/L12 during translocation as revealed by cryo-EM. , 2005, Molecular cell.

[9]  F. Hou,et al.  Ribosomal protein L10a, a bridge between trichosanthin and the ribosome. , 2005, Biochemical and biophysical research communications.

[10]  P. Shaw,et al.  Recent advances in trichosanthin, a ribosome-inactivating protein with multiple pharmacological properties. , 2005, Toxicon : official journal of the International Society on Toxinology.

[11]  Kevin Cowtan,et al.  research papers Acta Crystallographica Section D Biological , 2005 .

[12]  Y. Endo,et al.  Ribosomal Proteins at the Stalk Region Modulate Functional rRNA Structures in the GTPase Center* , 2002, The Journal of Biological Chemistry.

[13]  M. Tchórzewski.  The acidic ribosomal P proteins. , 2002, The international journal of biochemistry & cell biology.

[14]  P. Shaw,et al.  Trichosanthin interacts with acidic ribosomal proteins P0 and P1 and mitotic checkpoint protein MAD2B. , 2001, European journal of biochemistry.

[15]  L. Federici,et al.  The crystal structure of saporin SO6 from Saponaria officinalis and its interaction with the ribosome , 2000, FEBS letters.

[16]  I. Kurinov,et al.  X‐ray crystallographic analysis of the structural basis for the interaction of pokeweed antiviral protein with guanine residues of ribosomal RNA , 2008, Protein science : a publication of the Protein Society.

[17]  P. Gonzalo,et al.  Interaction of elongation factor eEF-2 with ribosomal P proteins. , 1999, European journal of biochemistry.

[18]  D E McRee,et al.  XtalView/Xfit--A versatile program for manipulating atomic coordinates and electron density. , 1999, Journal of structural biology.

[19]  J. Dinman,et al.  Pokeweed Antiviral Protein Accesses Ribosomes by Binding to L3* , 1999, The Journal of Biological Chemistry.

[20]  R J Read,et al.  Crystallography & NMR system: A new software suite for macromolecular structure determination. , 1998, Acta crystallographica. Section D, Biological crystallography.

[21]  A. Vagin,et al.  MOLREP: an Automated Program for Molecular Replacement , 1997 .

[22]  L Holm,et al.  Alignment of three-dimensional protein structures: network server for database searching. , 1996, Methods in enzymology.

[23]  C. Vater,et al.  Ricin A Chain Can Be Chemically Cross-linked to the Mammalian Ribosomal Proteins L9 and L10e (*) , 1995, The Journal of Biological Chemistry.

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

[25]  M. James,et al.  Crystal structure of the holotoxin from Shigella dysenteriae at 2.5 A resolution. , 1994, Nature structural biology.

[26]  M. James,et al.  Crystal structure of the holotoxino from Shigella dysenteriae at 2.5 Å resolution , 1994, Nature Structural Biology.

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

[28]  A. Monzingo,et al.  Structure of recombinant ricin A chain at 2.3 Å , 1993, Protein science : a publication of the Protein Society.

[29]  M. Brunori,et al.  A ribosomal protein is specifically recognized by saporin, a plant toxin which inhibits protein synthesis , 1992, FEBS letters.