The Amino-terminal Domain of Heat Shock Protein 90 (hsp90) That Binds Geldanamycin Is an ATP/ADP Switch Domain That Regulates hsp90 Conformation*

Many functions of the chaperone, heat shock protein 90 (hsp90), are inhibited by the drug geldanamycin that specifically binds hsp90. We have studied an amino-terminal domain of hsp90 whose crystal structure has recently been solved and determined to contain a geldanamycin-binding site. We demonstrate that, in solution, drug binding is exclusive to this domain. This domain also binds ATP linked to Sepharose through the γ-phosphate. Binding is specific for ATP and ADP and is inhibited by geldanamycin. Mutation of four glycine residues within two proposed ATP binding motifs diminishes both geldanamycin binding and the ATP-dependent conversion of hsp90 to a conformation capable of binding the co-chaperone p23. Since p23 binding requires regions outside the 1–221 domain of hsp90, these results indicate a common site for nucleotides and geldanamycin that regulates the conformation of other hsp90 domains.

[1]  C. Seeger,et al.  Hepadnavirus assembly and reverse transcription require a multi‐component chaperone complex which is incorporated into nucleocapsids , 1997, The EMBO journal.

[2]  R. Morimoto,et al.  The human cytosolic molecular chaperones hsp90, hsp70 (hsc70) and hdj‐1 have distinct roles in recognition of a non‐native protein and protein refolding. , 1996, The EMBO journal.

[3]  R. Jove,et al.  Raf exists in a native heterocomplex with hsp90 and p50 that can be reconstituted in a cell-free system. , 1993, The Journal of biological chemistry.

[4]  D. Toft,et al.  Mutational analysis of hsp90 binding to the progesterone receptor. , 1993, The Journal of biological chemistry.

[5]  R. Davis,et al.  The native structure of the activated Raf protein kinase is a membrane-bound multi-subunit complex. , 1994, The Journal of biological chemistry.

[6]  C. Kahn,et al.  The 90-kDa heat shock protein (hsp-90) possesses an ATP binding site and autophosphorylating activity. , 1991, The Journal of biological chemistry.

[7]  W. Pratt,et al.  The 23-kDa Acidic Protein in Reticulocyte Lysate Is the Weakly Bound Component of the hsp Foldosome That Is Required for Assembly of the Glucocorticoid Receptor into a Functional Heterocomplex with hsp90 (*) , 1995, The Journal of Biological Chemistry.

[8]  E. Alnemri,et al.  Nucleotides and Two Functional States of hsp90* , 1997, The Journal of Biological Chemistry.

[9]  R. Jove,et al.  The hsp90-binding Antibiotic Geldanamycin Decreases Raf Levels and Epidermal Growth Factor Signaling without Disrupting Formation of Signaling Complexes or Reducing the Specific Enzymatic Activity of Raf Kinase* , 1997, The Journal of Biological Chemistry.

[10]  L. Neckers,et al.  Depletion of p185erbB2, Raf-1 and mutant p53 proteins by geldanamycin derivatives correlates with antiproliferative activity , 1997, Cancer Chemotherapy and Pharmacology.

[11]  V. Thulasiraman,et al.  Effect of geldanamycin on the kinetics of chaperone-mediated renaturation of firefly luciferase in rabbit reticulocyte lysate. , 1996, Biochemistry.

[12]  C. Nicchitta,et al.  Interaction of Endoplasmic Reticulum Chaperone GRP94 with Peptide Substrates Is Adenine Nucleotide-independent* , 1997, The Journal of Biological Chemistry.

[13]  Susan S. Taylor,et al.  How do protein kinases discriminate between serine/threonine and tyrosine? Structural insights from the insulin receptor protein‐tyrosine kinase , 1995, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[14]  J. Brugge,et al.  Interaction of the Rous sarcoma virus protein pp60src with the cellular proteins pp50 and pp90. , 1986, Current topics in microbiology and immunology.

[15]  S. Ho,et al.  Site-directed mutagenesis by overlap extension using the polymerase chain reaction. , 1989, Gene.

[16]  J. Zheng,et al.  Structure of a peptide inhibitor bound to the catalytic subunit of cyclic adenosine monophosphate-dependent protein kinase. , 1991, Science.

[17]  J. Johnson,et al.  Characterization of a novel 23-kilodalton protein of unactive progesterone receptor complexes , 1994, Molecular and cellular biology.

[18]  Mikhail V. Blagosklonny,et al.  Disruption of the Raf-1-Hsp90 Molecular Complex Results in Destabilization of Raf-1 and Loss of Raf-1-Ras Association (*) , 1995, The Journal of Biological Chemistry.

[19]  L. Neckers,et al.  Destabilization of Raf-1 by geldanamycin leads to disruption of the Raf-1-MEK-mitogen-activated protein kinase signalling pathway , 1996, Molecular and cellular biology.

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

[21]  E. Alnemri,et al.  Cooperative action of Hsp70, Hsp90, and DnaJ proteins in protein renaturation. , 1996, Biochemistry.

[22]  L. Neckers,et al.  Inhibition of heat shock protein HSP90-pp60v-src heteroprotein complex formation by benzoquinone ansamycins: essential role for stress proteins in oncogenic transformation. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[23]  D. Wigley,et al.  Crystal structure of an N-terminal fragment of the DNA gyrase B protein , 1991, Nature.

[24]  O. Bischof,et al.  Cell cycle‐dependent association of Gag‐Mil and hsp90 , 1994, FEBS letters.

[25]  T. Smithgall,et al.  A pathway of multi-chaperone interactions common to diverse regulatory proteins: estrogen receptor, Fes tyrosine kinase, heat shock transcription factor Hsf1, and the aryl hydrocarbon receptor. , 1996, Cell stress & chaperones.

[26]  C. Kahn,et al.  ATP induces a conformational change of the 90-kDa heat shock protein (hsp90). , 1993, The Journal of biological chemistry.

[27]  L. Neckers,et al.  Mutant conformation of p53 translated in vitro or in vivo requires functional HSP90. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[28]  A. Nicolas,et al.  An atypical topoisomerase II from archaea with implications for meiotic recombination , 1997, Nature.

[29]  P. Russell,et al.  A role for Hsp90 in cell cycle control: Wee1 tyrosine kinase activity requires interaction with Hsp90. , 1994, The EMBO journal.

[30]  K. Yamamoto,et al.  A role for Hsp90 in retinoid receptor signal transduction. , 1995, Molecular biology of the cell.

[31]  F. Collins,et al.  Principles of Biochemistry , 1937, The Indian Medical Gazette.

[32]  L. Neckers,et al.  Geldanamycin selectively destabilizes and conformationally alters mutated p53. , 1995, Oncogene.

[33]  R. Rimerman,et al.  Progesterone receptor structure and function altered by geldanamycin, an hsp90-binding agent , 1995, Molecular and cellular biology.

[34]  Y. Takahashi,et al.  Characterization of the hydrophobic region of heat shock protein 90. , 1991, Journal of biochemistry.

[35]  T. Haystead,et al.  Gamma-phosphate-linked ATP-sepharose for the affinity purification of protein kinases. Rapid purification to homogeneity of skeletal muscle mitogen-activated protein kinase kinase. , 1993, European journal of biochemistry.

[36]  F. Hartl,et al.  Pharmacologic shifting of a balance between protein refolding and degradation mediated by Hsp90. , 1996, Proceedings of the National Academy of Sciences of the United States of America.

[37]  P Bork,et al.  Positionally cloned human disease genes: patterns of evolutionary conservation and functional motifs. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[38]  C. Walsh,et al.  Hsp90 chaperonins possess ATPase activity and bind heat shock transcription factors and peptidyl prolyl isomerases. , 1993, The Journal of biological chemistry.

[39]  Laurence H. Pearl,et al.  A molecular clamp in the crystal structure of the N-terminal domain of the yeast Hsp90 chaperone , 1997, Nature Structural Biology.

[40]  D. J. Barrett,et al.  Hsp90-mediated folding of the lymphoid cell kinase p56lck. , 1996, Biochemistry.

[41]  R. Kolodner,et al.  Biochemistry and genetics of eukaryotic mismatch repair. , 1996, Genes & development.

[42]  R. Schnur,et al.  Preparation of 17-amino-22-(4′-azido-3′-125iodophenacyl)-17-demethoxygeldanamycin (1): An ansamycin for photoaffinity labeling , 1994 .

[43]  J. Johnson,et al.  A novel chaperone complex for steroid receptors involving heat shock proteins, immunophilins, and p23. , 1994, The Journal of biological chemistry.

[44]  E. Alnemri,et al.  The steroid binding domain influences intracellular solubility of the baculovirus overexpressed glucocorticoid and mineralocorticoid receptors. , 1993, Biochemistry.

[45]  L. Poellinger,et al.  The Basic Helix-Loop-Helix/PAS Factor Sim Is Associated with hsp90: , 1996, The Journal of Biological Chemistry.

[46]  R. Matts,et al.  Association of Hsp90 with cellular Src-family kinases in a cell-free system correlates with altered kinase structure and function. , 1994, Biochemistry.

[47]  J. Buchner,et al.  Transient Interaction of Hsp90 with Early Unfolding Intermediates of Citrate Synthase , 1995, The Journal of Biological Chemistry.

[48]  W. Kolch,et al.  Nerve Growth Factor-mediated Activation of the Mitogen-activated Protein (MAP) Kinase Cascade Involves a Signaling Complex Containing B-Raf and HSP90* , 1996, The Journal of Biological Chemistry.

[49]  S A Benner,et al.  A predicted consensus structure for the N‐Terminal fragment of the heat shock protein HSP90 family , 1997, Proteins.

[50]  J. Reinstein,et al.  Assessment of the ATP Binding Properties of Hsp90 (*) , 1996, The Journal of Biological Chemistry.

[51]  J. Harper,et al.  Mammalian p50Cdc37 is a protein kinase-targeting subunit of Hsp90 that binds and stabilizes Cdk4. , 1996, Genes & development.

[52]  C. Seeger,et al.  Hsp90 is required for the activity of a hepatitis B virus reverse transcriptase. , 1996, Proceedings of the National Academy of Sciences of the United States of America.