Details
| Original language | English |
|---|---|
| Pages (from-to) | 562–574 |
| Number of pages | 13 |
| Journal | CHEMBIOCHEM |
| Volume | 19 |
| Issue number | 6 |
| Early online date | 18 Dec 2017 |
| Publication status | Published - 25 Mar 2018 |
Abstract
Thirteen new reblastatin derivatives, with alkynyl, amino and fluoro substituents on the aromatic ring, were prepared by a chemo-biosynthetic approach using an AHBA(−) mutant strain of Streptomyces hygroscopicus, the geldanamycin producer. The inhibitory potencies of these mutaproducts and of an extended library of natural products and derivatives were probed with purified heat shock proteins (Hsps), obtained from Leishmania braziliensis (LbHsp90) as well as from human sources (HsHsp90). We determined the activities of potential inhibitors by means of a displacement assay in which fluorescence-labelled ATP competes for the ATP binding sites of Hsps in the presence of the inhibitor in question. The results were compared with those of cell-based assays and, in selected cases, of isothermal titration calorimetry (ITC) measurements. In essence, reblastatin derivatives are also able to bind effectively to the ATP-binding site of LbHsp90, and for selected derivatives, moderate differences in binding to LbHsp90 and HsHsp90 were encountered. This work demonstrates that parasitic heat shock proteins can be developed as potential pharmaceutical targets.
Keywords
- geldanamycin, heat shock proteins, inhibitors, Leishmania, microarrays
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Biochemistry
- Biochemistry, Genetics and Molecular Biology(all)
- Molecular Medicine
- Biochemistry, Genetics and Molecular Biology(all)
- Molecular Biology
- Chemistry(all)
- Organic Chemistry
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In: CHEMBIOCHEM, Vol. 19, No. 6, 25.03.2018, p. 562–574.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Heat Shock Proteins Revisited:
T2 - Using a Mutasynthetically Generated Reblastatin Library to Compare the Inhibition of Human and Leishmania Hsp90s
AU - Mohammadi-Ostad-Kalayeh, Sona
AU - Stahl, Frank
AU - Scheper, Thomas
AU - Kock, Klaus
AU - Herrmann, Christian
AU - Heleno Batista, Fernanda Aparecida
AU - Borges, Júlio César
AU - Sasse, Florenz
AU - Eichner, Simone
AU - Ongouta, Jekaterina
AU - Zeilinger, Carsten
AU - Kirschning, Andreas
N1 - Funding information: This work was supported by the Deutsche Forschungsgemein-schaft (DFG, grant Ki 13-1). J.C.B. thanks the S¼o Paulo Research Foundation (FAPESP 2012/50161-8 and 2014/07206-6) and the Brazilian Council for Scientific and Technological Development (CNPq 303129/2015-8 and 471415/2013-8) for financial support.
PY - 2018/3/25
Y1 - 2018/3/25
N2 - Thirteen new reblastatin derivatives, with alkynyl, amino and fluoro substituents on the aromatic ring, were prepared by a chemo-biosynthetic approach using an AHBA(−) mutant strain of Streptomyces hygroscopicus, the geldanamycin producer. The inhibitory potencies of these mutaproducts and of an extended library of natural products and derivatives were probed with purified heat shock proteins (Hsps), obtained from Leishmania braziliensis (LbHsp90) as well as from human sources (HsHsp90). We determined the activities of potential inhibitors by means of a displacement assay in which fluorescence-labelled ATP competes for the ATP binding sites of Hsps in the presence of the inhibitor in question. The results were compared with those of cell-based assays and, in selected cases, of isothermal titration calorimetry (ITC) measurements. In essence, reblastatin derivatives are also able to bind effectively to the ATP-binding site of LbHsp90, and for selected derivatives, moderate differences in binding to LbHsp90 and HsHsp90 were encountered. This work demonstrates that parasitic heat shock proteins can be developed as potential pharmaceutical targets.
AB - Thirteen new reblastatin derivatives, with alkynyl, amino and fluoro substituents on the aromatic ring, were prepared by a chemo-biosynthetic approach using an AHBA(−) mutant strain of Streptomyces hygroscopicus, the geldanamycin producer. The inhibitory potencies of these mutaproducts and of an extended library of natural products and derivatives were probed with purified heat shock proteins (Hsps), obtained from Leishmania braziliensis (LbHsp90) as well as from human sources (HsHsp90). We determined the activities of potential inhibitors by means of a displacement assay in which fluorescence-labelled ATP competes for the ATP binding sites of Hsps in the presence of the inhibitor in question. The results were compared with those of cell-based assays and, in selected cases, of isothermal titration calorimetry (ITC) measurements. In essence, reblastatin derivatives are also able to bind effectively to the ATP-binding site of LbHsp90, and for selected derivatives, moderate differences in binding to LbHsp90 and HsHsp90 were encountered. This work demonstrates that parasitic heat shock proteins can be developed as potential pharmaceutical targets.
KW - geldanamycin
KW - heat shock proteins
KW - inhibitors
KW - Leishmania
KW - microarrays
UR - http://www.scopus.com/inward/record.url?scp=85042103305&partnerID=8YFLogxK
U2 - 10.1002/cbic.201700616
DO - 10.1002/cbic.201700616
M3 - Article
C2 - 29265716
AN - SCOPUS:85042103305
VL - 19
SP - 562
EP - 574
JO - CHEMBIOCHEM
JF - CHEMBIOCHEM
SN - 1439-4227
IS - 6
ER -