Native Size-Exclusion Chromatography–Based Mass Spectrometry Reveals New Components of the Early Heat Shock Protein 90 Inhibition Response Among Limited Global Changes

Samant, Rahul S.; Batista, Sílvia; Larance, Mark; Özer, Buğra; Milton, Christopher; Bludau, Isabell; Wu, Estelle; Biggins, Laura; Andrews, Simon; Hervieu, Alexia; Johnston, Harvey E.; Al‐Lazikani, Bissan; Lamond, Angus I.; Clarke, Paul; Workman, Paul

doi:10.1016/j.mcpro.2022.100485

Cited by 3 publications

(5 citation statements)

References 130 publications

(190 reference statements)

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“…Over the years, various methods have been used to characterize Hsp interactomes with an emphasis on cataloging their client proteins in efforts to understand substrate recognition. These methods include affinity capture (antibody‐ and small molecule‐based) [44–49], in situ protein proximity labeling [50,51], and co‐fractionation [52]. Due to the transient nature of chaperone complexes, none of these methods will likely provide an entire picture of the interactome [53].…”

Section: Chaperone Interactomes and Insights Into Substrate Recognitionmentioning

confidence: 99%

“…Recent studies have estimated that more than 20% of the human proteome and over 60% of the kinome interacts with Hsp90 and can be largely classified as either co‐chaperones or client proteins [44,45,52,64]. Global analysis of the Hsp90 interactome provided insight that specific co‐chaperones can drive the recruitment of various protein classes [44].…”

Section: Chaperone Interactomes and Insights Into Substrate Recognitionmentioning

confidence: 99%

“…TRAP1 is reported to interact with two of the three dehydrogenases in the TCA cycle – mitochondrial isocitrate dehydrogenase (IDH3) [52] and succinate dehydrogenase (SDH) [116] (Fig. 3A).…”

Section: Supramolecular Metabolic Enzyme Assembliesmentioning

confidence: 99%

“…Using native size‐exclusion chromatography, two of the three subunits were shown to disengage from the complex upon Hsp90 inhibition with 17‐AAG. This caused a greater than 50% reduction in IDH3 enzymatic activity in HT29, HCT116, and BT474 cancer cell lines [52]. Other IDH complexes (IDH1 and IDH2) that utilize NADP + as a cofactor did not show the same reliance on TRAP1 activity.…”

Section: Supramolecular Metabolic Enzyme Assembliesmentioning

confidence: 99%

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The roles of molecular chaperones in regulating cell metabolism

Binder

Pedley

2023

FEBS Letters

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Fluctuations in nutrient and biomass availability, often as a result of disease, impart metabolic challenges that must be overcome in order to sustain cell survival and promote proliferation. Cells adapt to these environmental changes and stresses by adjusting their metabolic networks through a series of regulatory mechanisms. Our understanding of these rewiring events has largely been focused on those genetic transformations that alter protein expression and the biochemical mechanisms that change protein behavior, such as post‐translational modifications and metabolite‐based allosteric modulators. Mounting evidence suggests that a class of proteome surveillance proteins called molecular chaperones also can influence metabolic processes. Here, we summarize several ways the Hsp90 and Hsp70 chaperone families act on human metabolic enzymes and their supramolecular assemblies to change enzymatic activities and metabolite flux. We further highlight how these chaperones can assist in the translocation and degradation of metabolic enzymes. Collectively, these studies provide a new view for how metabolic processes are regulated to meet cellular demand and inspire new avenues for therapeutic intervention.

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Section: Chaperone Interactomes and Insights Into Substrate Recognitionmentioning

confidence: 99%

Section: Chaperone Interactomes and Insights Into Substrate Recognitionmentioning

confidence: 99%

Section: Supramolecular Metabolic Enzyme Assembliesmentioning

confidence: 99%

Section: Supramolecular Metabolic Enzyme Assembliesmentioning

confidence: 99%

See 2 more Smart Citations

The roles of molecular chaperones in regulating cell metabolism

Binder

Pedley

2023

FEBS Letters

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“…This modeling adequately describes the dissemination over time of newly introduced compounds in animals and is most often used in preclinical research [ 38 , 42 ]. The use of sensitive and robust analytical techniques, such as high-performance liquid chromatographic (HPLC), has contributed to the advancement of clinical pharmacokinetics, especially in the development of methods for the determination of Tanespimycin (17-AAG) in biological fluids [ 43 , 44 , 45 ].…”

Section: Introductionmentioning

confidence: 99%

Pharmacokinetics, Dose-Proportionality, and Tolerability of Intravenous Tanespimycin (17-AAG) in Single and Multiple Doses in Dogs: A Potential Novel Treatment for Canine Visceral Leishmaniasis

Ferrante,

Leite,

Fontes

et al. 2024

Pharmaceuticals

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In the New World, dogs are considered the main reservoir of visceral leishmaniasis (VL). Due to inefficacies in existing treatments and the lack of an efficient vaccine, dog culling is one of the main strategies used to control disease, making the development of new therapeutic interventions mandatory. We previously showed that Tanespimycin (17-AAG), a Hsp90 inhibitor, demonstrated potential for use in leishmaniasis treatment. The present study aimed to test the safety of 17-AAG in dogs by evaluating plasma pharmacokinetics, dose-proportionality, and the tolerability of 17-AAG in response to a dose-escalation protocol and multiple administrations at a single dose in healthy dogs. Two protocols were used: Study A: four dogs received variable intravenous (IV) doses (50, 100, 150, 200, or 250 mg/m2) of 17-AAG or a placebo (n = 4/dose level), using a cross-over design with a 7-day “wash-out” period; Study B: nine dogs received three IV doses of 150 mg/m2 of 17-AAG administered at 48 h intervals. 17-AAG concentrations were determined by a validated high-performance liquid chromatographic (HPLC) method: linearity (R2 = 0.9964), intra-day precision with a coefficient of variation (CV) ≤ 8%, inter-day precision (CV ≤ 20%), and detection and quantification limits of 12.5 and 25 ng/mL, respectively. In Study A, 17-AAG was generally well tolerated. However, increased levels of liver enzymes–alanine aminotransferase (ALT), aspartate aminotransferase (AST), and gamma-glutamyl transferase (GGT)–and bloody diarrhea were observed in all four dogs receiving the highest dosage of 250 mg/m2. After single doses of 17-AAG (50–250 mg/m2), maximum plasma concentrations (Cmax) ranged between 1405 ± 686 and 9439 ± 991 ng/mL, and the area under the curve (AUC) plotting plasma concentration against time ranged between 1483 ± 694 and 11,902 ± 1962 AUC 0–8 h μg/mL × h, respectively. Cmax and AUC parameters were dose-proportionate between the 50 and 200 mg/m2 doses. Regarding Study B, 17-AAG was found to be well tolerated at multiple doses of 150 mg/m2. Increased levels of liver enzymes–ALT (28.57 ± 4.29 to 173.33 ± 49.56 U/L), AST (27.85 ± 3.80 to 248.20 ± 85.80 U/L), and GGT (1.60 ± 0.06 to 12.70 ± 0.50 U/L)–and bloody diarrhea were observed in only 3/9 of these dogs. After the administration of multiple doses, Cmax and AUC 0–48 h were 5254 ± 2784 μg/mL and 6850 ± 469 μg/mL × h in plasma and 736 ± 294 μg/mL and 7382 ± 1357 μg/mL × h in tissue transudate, respectively. In conclusion, our results demonstrate the potential of 17-AAG in the treatment of CVL, using a regimen of three doses at 150 mg/m2, since it presents the maintenance of high concentrations in subcutaneous interstitial fluid, low toxicity, and reversible hepatotoxicity.

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