Crystal structure and molecular modeling of 17-DMAG in complex with human Hsp90.

Hsp90 is an attractive chemotherapeutic target because it chaperones the folding of proteins found in multiple signal transduction pathways. We describe the 1.75 A resolution crystal structure of human Hsp90 alpha (residues 9-236) complexed with 17-desmethoxy-17-N,N-dimethylaminoethylamino-geldanamycin (17-DMAG). The structure revealed an altered set of interactions between the 17-substituent and the protein compared to geldanamycin and the 17-dimethylaminoethyl moiety pointing into solvent, but otherwise was similar to that reported for the complex with geldanamycin. Targeted molecular dynamics simulations and energetic analysis indicate that geldanamycin undergoes two major conformational changes when it binds Hsp90, with the key step of the conversion being the trans to cis conformational change of the macrocycle amide bond. We speculate that 17-DMAG analogs constrained to a cis-amide in the ground state could provide a significant increase in affinity for Hsp90.

[1]  P. Kollman,et al.  A well-behaved electrostatic potential-based method using charge restraints for deriving atomic char , 1993 .

[2]  B. Segnitz,et al.  The Function of Steroid Hormone Receptors Is Inhibited by the hsp90-specific Compound Geldanamycin* , 1997, The Journal of Biological Chemistry.

[3]  P. Miller,et al.  erbB-2 oncogene inhibition by geldanamycin derivatives: synthesis, mechanism of action, and structure-activity relationships. , 1995, Journal of medicinal chemistry.

[4]  C. Cramer,et al.  An SCF Solvation Model for the Hydrophobic Effect and Absolute Free Energies of Aqueous Solvation , 1992, Science.

[5]  M. L. Connolly.  Analytical molecular surface calculation , 1983 .

[6]  P. W. Bowman,et al.  PHS Public Health Service , 1963 .

[7]  K D Cowtan,et al.  Improvement of macromolecular electron-density maps by the simultaneous application of real and reciprocal space constraints. , 1993, Acta crystallographica. Section D, Biological crystallography.

[8]  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.

[9]  M Karplus,et al.  Molecular switch in signal transduction: reaction paths of the conformational changes in ras p21. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[10]  L. Neckers,et al.  p185erbB2 binds to GRP94 in vivo. Dissociation of the p185erbB2/GRP94 heterocomplex by benzoquinone ansamycins precedes depletion of p185erbB2. , 1996, The Journal of biological chemistry.

[11]  M. Egorin,et al.  Pharmacokinetics, tissue distribution, and metabolism of 17-(dimethylaminoethylamino)-17-demethoxygeldanamycin (NSC 707545) in CD2F1 mice and Fischer 344 rats , 2001, Cancer Chemotherapy and Pharmacology.

[12]  B. Honig,et al.  Classical electrostatics in biology and chemistry. , 1995, Science.

[13]  J. Greenstein,et al.  Chemistry of the Amino Acids , 1961 .

[14]  L. Pearl,et al.  Structural basis for inhibition of the Hsp90 molecular chaperone by the antitumor antibiotics radicicol and geldanamycin. , 1999, Journal of medicinal chemistry.

[15]  Yufeng Shen,et al.  Conformational pathways in the gating of Escherichia coli mechanosensitive channel , 2002, Proceedings of the National Academy of Sciences of the United States of America.

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

[17]  D. Case,et al.  Theory and applications of the generalized born solvation model in macromolecular simulations , 2000, Biopolymers.

[18]  M Karplus,et al.  A Dynamic Model for the Allosteric Mechanism of GroEL , 2000 .

[19]  M. Grever,et al.  Preclinical pharmacologic evaluation of geldanamycin as an antitumor agent , 2004, Cancer Chemotherapy and Pharmacology.

[20]  R. Schnur,et al.  Tandem [3,3]-sigmatropic rearrangements in an ansamycin : stereospecific conversion of an (S)-allylic alcohol to an (S)-allylic amine derivative , 1994 .

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

[22]  D. Mottet,et al.  Hypoxia‐induced activation of HIF‐1: role of HIF‐1α‐Hsp90 interaction , 1999 .

[23]  C. Diorio,et al.  Inhibition of the oncogene product p185erbB-2 in vitro and in vivo by geldanamycin and dihydrogeldanamycin derivatives. , 1995, Journal of medicinal chemistry.

[24]  G. Ciccotti,et al.  Numerical Integration of the Cartesian Equations of Motion of a System with Constraints: Molecular Dynamics of n-Alkanes , 1977 .

[25]  A G Leslie,et al.  Biological Crystallography Integration of Macromolecular Diffraction Data , 2022 .

[26]  P. Kollman,et al.  Calculating structures and free energies of complex molecules: combining molecular mechanics and continuum models. , 2000, Accounts of chemical research.

[27]  D. Case,et al.  Molecular Dynamics Simulations of Nucleic Acids with a Generalized Born Solvation Model , 2000 .

[28]  Conrad C. Huang,et al.  The MIDAS display system , 1988 .

[29]  P. Kollman,et al.  A Second Generation Force Field for the Simulation of Proteins, Nucleic Acids, and Organic Molecules , 1995 .

[30]  L. Weber,et al.  Sequence and regulation of a gene encoding a human 89-kilodalton heat shock protein , 1989, Molecular and cellular biology.

[31]  Neal Rosen,et al.  Crystal Structure of an Hsp90–Geldanamycin Complex: Targeting of a Protein Chaperone by an Antitumor Agent , 1997, Cell.

[32]  K. Rinehart,et al.  Chemistry of the ansamycin antibiotics. , 1976, Fortschritte der Chemie organischer Naturstoffe = Progress in the chemistry of organic natural products. Progres dans la chimie des substances organiques naturelles.

[33]  P. Krüger,et al.  Targeted molecular dynamics: a new approach for searching pathways of conformational transitions. , 1994, Journal of molecular graphics.

[34]  P. Kraulis.  A program to produce both detailed and schematic plots of protein structures , 1991 .