Biochemical and Biophysical Research Communications
The induction mechanism of the molecular chaperone HSP70 in the gastric mucosa by Geranylgeranylacetone (HSP-inducer)
Section snippets
Plasmid or phagemid construction
A full-length human HSP70 cDNA was kindly provided by Dr. Richard Morimoto, Northwestern University (Evanston, Ill, U.S.A.). The polymerase chain reaction-amplified BamHI–HindIII fragments encoding the cDNA sequence, the N-terminal domain of HSP70 (residues 1–380) or the C-terminal domain of HSP70 (residues 381–640), were inserted into the digested pQE31 plasmid vector (Quiagen, Hilden, Germany). The N- or C-terminal domain of HSP70 was overexpressed in Escherichia coli M15 [pREP4] cells as an
GGA-affinity column chromatography
We investigated GGA-binding proteins using a GGA-affinity column. In general, the affinity column chromatography is defined as follows: a purified molecule is reversibly adsorbed on the ligand which is fixed to the matrix [25], [26]. To avoid nonspecific binding proteins, GGA-affinity columns were thoroughly washed with conditioning buffer. Proteins were eluted with 10 mM GGA from GGA-affinity column or control mock column, and eluants were detected on SDS/PAGE. Although no protein bands were
Discussion
GGA, an anti-ulcer drug, was first introduced as a nontoxic inducer of molecular chaperone HSP70 in rat gastric mucosa [14]. However, the induction mechanisms of HSP70 by GGA remain to be elucidated. In the present study, to address the questions, we investigated the specific GGA-binding proteins using a GGA-affinity column. The specific binding protein with a 70-kDa molecular mass was constitutively expressed molecular chaperone HSP70 (known as HSP73). The purified wild-type HSP70 also showed
Acknowledgments
This work was supported in part by a Grant-in-Aid for Scientific Research to H.I. (Exploratory Research: No. 16651056) and M.O. (No. 17590610) from the Japanese Ministry of Education, Science, Sports and Culture.
References (30)
- et al.
The Hsp70 and Hsp60 chaperone machines
Cell
(1998) - et al.
The ATPase core of a clathrin uncoating protein
J. Biol. Chem.
(1987) - et al.
Effect of nucleotide on the binding of peptides to 70-kDa heat shock protein
J. Biol. Chem.
(1995) - et al.
Geranylgeranylacetone induces heat shock proteins in cultured guinea pig gastric mucosal cells and rat gastric mucosa
Gastroenterology
(1996) - et al.
Effects of geranyl-geranyl-acetone administration before heat shock preconditioning for conferring tolerance against ischemia-reperfusion injury in rat livers
J. Lab. Clin. Med.
(2000) - et al.
Astroglial activation accompanies heat shock protein upregulation in rat brain following single oral dose of geranylgeranylacetone
Brain. Res.
(2003) - et al.
73-kDa molecular chaperone HSP73 is a direct target of antibiotic gentamicin
J. Biol. Chem.
(2004) - et al.
Affinity chromatography of serum albumin with fatty acids immobilized on agarose
J. Biol. Chem.
(1973) - et al.
Identification of the peptide binding domain of hsc70. 18-Kilodalton fragment located immediately after ATPase domain is sufficient for high affinity binding
J. Biol. Chem.
(1993) - et al.
Hop as an adaptor in the heat shock protein 70 (Hsp70) and hsp90 chaperone machinery
J. Biol. Chem.
(1998)
Pathways of chaperone-mediated protein folding in the cytosol
Nat. Rev. Mol. Cell. Biol.
The stress (heat shock) proteins
Int. J. Biochem.
The human cytosolic molecular chaperones hsp90, hsp70 (hsc70) and hdj-1 have distinct roles in recognition of a non-native protein and protein refolding
EMBO J.
Three-dimensional structure of the ATPase fragment of a 70 K heat-shock cognate protein
Nature.
Kinetics of molecular chaperone action
Science.
Cited by (60)
Targeting Hsp70: A possible therapy for cancer
2016, Cancer LettersAnthocyanin-rich tea Sunrouge upregulates expressions of heat shock proteins in the gastrointestinal tract of ICR mice: A comparison with the conventional tea cultivar Yabukita
2015, Journal of Food and Drug AnalysisCitation Excerpt :Geldanamycin, a natural benzoquinone antibiotic, has been shown to inhibit HSP90 by binding to the adenosine diphosphate-adenosine triphosphate-binding domain for upregulating many hsp genes [29]. Geranylgeranylacetone, however, has also been described as a potent antiulcer drug, which is capable of increasing the expression of inducible HSPs via binding to the C-terminal of HSP70 [30], in experimental rodents [31] and healthy volunteers [32]. In addition, Ahmed et al [33] have recently screened 80 compounds from medicinal plants for their HSP70 inducing activities in U937 human lymphoma cells.
The novolactone natural product disrupts the allosteric regulation of Hsp70
2015, Chemistry and Biology