Molecular determinants of Hsp90 dependence of Src kinase revealed by deep mutational scanning

Nguyen, Vanessa P.; Ahler, Ethan; Sitko, Katherine A.; Stephany, Jason J.; Maly, Dustin J.; Fowler, Douglas M.

doi:10.1002/pro.4656

Cited by 6 publications

(1 citation statement)

References 60 publications

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“…HSF1 is a transcription factor that can induce the expression of HSPs (Kurop et al, 2021). Under non-stress conditions, HSF1 exists in an inactive monomer form and forms a complex with HSP90 (Nguyen et al, 2023). Studies found that the heat shock response (HSR) is regulated by the phosphorylation of HSF1 and the dissociation of HSP90 (Gong et al, 2019).…”

Section: Discussionmentioning

confidence: 99%

Kaempferol exerts antioxidant effects in age‐related diminished ovarian reserve by regulating the HSP90/NRF2 pathway

Hua,

Zhang,

Han

et al. 2023

Chem Biol Drug Des

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Kaempferol is the active ingredient of Er‐Xian decoction (EXD), a traditional Chinese medicine formula used clinically to treat ovarian dysfunction, but the mechanism of kaempferol relieving age‐related diminished ovarian reserve (AR‐DOR) is still unclear. In this study, 36 volunteers and 78 DOR patients (37 patients with EXD treatment) were enrolled in the clinical research. Meanwhile, 32‐week‐old female mice were used to establish the AR‐DOR model, and these model mice were intragastrically administered with 100 mg/kg kaempferol in the presence or absence of 200 mg/kg geranylgeranylacetone (GGA) or 1 mg/kg geldanamycin (GDA). The effects of kaempferol on serum hormone levels and oxidative stress‐related indexes were detected by enzyme‐linked immunosorbent assay. Antral follicle count (AFC) was determined by hematoxylin–eosin staining. The protein levels of HSP90 and nuclear factor erythroid 2‐related factor 2 (NRF2) were assayed by Western blot. This study displayed that the serum anti‐Mullerian hormone (AMH) level in DOR patients with EXD treatment was higher than that in DOR patients without EXD treatment. Kaempferol treatment reversed the low levels of AMH, estradiol (E2), AFC, superoxide dismutase (SOD), and catalase (CAT), as well as the high levels of follicle‐stimulating hormone (FSH), reactive oxygen species (ROS), and malonaldehyde (MDA). The results showed that HSP90 was predicted to have high affinity with kaempferol, and its expression was inhibited by kaempferol, while the expression of NRF2, the target of HSP90, was up‐regulated by kaempferol. However, the above effects of kaempferol were reversed by GGA. On the contrary, GDA enhanced the therapeutic effects of kaempferol on AR‐DOR mice. Moreover, the treatment of kaempferol resulted in a reduction in the phosphorylation level of heat shock factor 1 (HSF1), the transcription factor associated with HSP90, and an increase in the phosphorylation level of Src, a client protein of HSP90. In summary, kaempferol exerts an antioxidant effect on AR‐DOR by inhibiting HSP90 expression to up‐regulate NRF2 expression. This study provides a theoretical basis for the clinical application of kaempferol in AR‐DOR.

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Section: Discussionmentioning

confidence: 99%

Kaempferol exerts antioxidant effects in age‐related diminished ovarian reserve by regulating the HSP90/NRF2 pathway

Hua,

Zhang,

Han

et al. 2023

Chem Biol Drug Des

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Molecular determinants of Hsp90 dependence of Src kinase revealed by deep mutational scanning

et al. 2023

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Hsp90 is a molecular chaperone involved in the refolding and activation of numerous protein substrates referred to as clients. While the molecular determinants of Hsp90 client specificity are poorly understood and limited to a handful of client proteins, strong clients are thought to be destabilized and conformationally extended. Here, we measured the phosphotransferase activity of 3929 variants of the tyrosine kinase Src in both the presence and absence of an Hsp90 inhibitor. We identified 84 previously unknown functionally dependent client variants. Unexpectedly, many destabilized or extended variants were not functionally dependent on Hsp90. Instead, functionally dependent client variants were clustered in the αF pocket and β1-β2 strand regions of Src,which have yet to be described in driving Hsp90 dependence. Hsp90 dependence was also strongly correlated with kinase activity. We found that a combination of activation, global extension, and general conformational flexibility, primarily induced by variants at the αF pocket and β1-β2 strands, was necessary to render Src functionally dependent on Hsp90. Moreover, the degree of activation and flexibility required to transform Src into a functionally dependent client varied with variant location, suggesting that a combination of regulatory domain disengagement and catalytic domain flexibility are required for chaperone dependence. Thus, by studying the chaperone dependence of a massive number of variants, we highlight factors driving Hsp90 client specificity and propose a model of chaperone-kinase interactions.

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Profiling of drug resistance in Src kinase at scale uncovers a regulatory network coupling autoinhibition and catalytic domain dynamics

Chakraborty,

Ahler,

Simon

et al. 2024

Cell Chemical Biology

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Molecular determinants of Hsp90 dependence of Src kinase revealed by deep mutational scanning

Cited by 6 publications

References 60 publications

Kaempferol exerts antioxidant effects in age‐related diminished ovarian reserve by regulating the HSP90/NRF2 pathway

Kaempferol exerts antioxidant effects in age‐related diminished ovarian reserve by regulating the HSP90/NRF2 pathway

Molecular determinants of Hsp90 dependence of Src kinase revealed by deep mutational scanning

Profiling of drug resistance in Src kinase at scale uncovers a regulatory network coupling autoinhibition and catalytic domain dynamics

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