In vivo function of the molecular chaperone Hsp90 is ATP-dependent and requires the full-length protein. Our earlier studies predicted a second C-terminal ATP-binding site in Hsp90. By applying direct biochemical approaches, we mapped two ATP-binding sites and unveiled the C-terminal ATP-binding site as the first example of a cryptic chaperone nucleotide-binding site, which is opened by occupan...

Energetic coupling of two molecular events in a protein molecule is ubiquitous in biochemical reactions mediated by proteins, such as catalysis and signal transduction. Here, we investigate energetic coupling between ligand binding and folding of a dimer using a model system that shows three-state equilibrium unfolding of an exceptional quality. The homodimeric Escherichia coli cofactor-depende...

Bernhard Huchzermeyer Botanisches Institut, Tierärztliche Hochschule, Bünteweg 17d, D-3000 Hannover 71, Bundesrepublik Deutschland Z. Naturforsch. 43c, 213-218 (1988); received July 27/November 19, 1987 Chloroplast, Coupling Factor, Binding Sites, ATPase, Phosphorylation A single binding site for phosphate was found on isolated chloroplast coupling factor in the absence of nucleotides. In our e...

1. The fluorescent ATP analogue 1,N6-etheno-ATP is a good substrate and an efficient allosteric inhibitor of rabbit skeletal-muscle phosphofructokinase. 2. Fluorescence energy transfer occurs between bound 1,N6-etheno-ATP and phosphofructokinase. 1,N6-Etheno-ATP fluorescence is enhanced, intrinsic protein fluorescence is quenched, and the excitation spectrum of 1,N6-etheno-ATP fluorescence is c...

The 90-kDa heat shock protein (Hsp90) is a molecular chaperone that assists both in ATP-independent sequestration of damaged proteins, and in ATP-dependent folding of numerous targets, such as nuclear hormone receptors and protein kinases. Recent work from our lab and others has established the existence of a second, C-terminal nucleotide binding site besides the well characterized N-terminal, ...

CFTR, the chloride channel mutated in cystic fibrosis (CF) patients, is opened by ATP binding to two cytosolic nucleotide binding domains (NBDs), but pore-domain mutations may also impair gating. ATP-bound NBDs dimerize occluding two nucleotides at interfacial binding sites; one site hydrolyzes ATP, the other is inactive. The pore opens upon tightening, and closes upon disengagement, of the cat...

Type 1 ryanodine receptor (RyR1) releases Ca2+ ions from the sarcoplasmic reticulum of skeletal muscle to initiate contraction. Multiple endogenous and exogenous effectors activate RyR1. Cryo-electron microscopy has identified binding sites three co-activators Ca2+, ATP caffeine, however, mechanism co-regulation synergy between these activators remains be determined. Here, we tested hypothesis ...

The development of selective inhibitors of protein kinases is challenging because of the significant conservation of the ATP binding site. Here, we describe a new mechanism by which the protein kinase CK2α can be selectively inhibited using features outside the ATP site. We have identified a new binding site for small molecules on CK2α adjacent to the ATP site and behind the αD loop, termed the...

Abl kinase inhibitors targeting the ATP binding pocket are currently employed as potent anti-leukemogenic agents but drug resistance has become a significant clinical limitation. Recently, a compound that binds to the myristate pocket of Abl (GNF-5) was shown to act cooperatively with nilotinib, an ATP-competitive inhibitor to target the recalcitrant "T315I" gatekeeper mutant of Bcr-Abl. To unc...

Prostate cancer is hormone-dependent and the androgen receptor (AR) involved in its development. AR a transcription factor that activated by ligand binding, result translocation into nucleus, where it initiates gene transcription. In an inactive state cytoplasm exists as complex with heat shock protein 90 (HSP90) some other proteins. When agonist binds, conformational change occurs, resulting H...